JPS63501482A - Method and apparatus for separating and isolating blood or bone marrow components - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Separation of blood or bone marrow components Oyo Oyo 11 No IJ The present invention allows cells of blood-forming tissues such as bone marrow or blood to be separated by centrifugation. A method and apparatus for separating and isolating these components, comprising: A blood or blood vessel is connected to one or more receiving vessels through an outflow opening. or as a container for bone marrow.

Blood is made up of four components, and these components are classified in order of increasing specific gravity. These are plasma, platelets, red blood cells and white blood cells. In the case of red blood cells, older red blood cells There is no distinction between gerocytes, which are blood cells, and neocytes, which are newly formed blood cells. be done. The average lifespan of red blood cells is approximately 90 days. Therefore, "new" red blood cells are compared to can remain viable for a long time, which is very important in the case of blood transfusions! It's important Ru. The specific gravity of red blood cells increases over time and is therefore determined by centrifugation based on lifespan. It is possible to determine the distribution of red blood cells. White blood cells and platelets (“several layers” and total In total, about 1% of the normal blood volume. Also, the total volume Approximately 45% of this is made up of red blood cells, and after centrifugation, neocytes (new blood cells) Approximately 50% of the total volume resides within the red blood cell layer, which occupies approximately 10% of the total volume.

Conventionally, the components that make up the soft calendar and neosite were prepared according to known methods. and then isolated from each other. Regarding this, for example, .

One problem with this approach, see European Patent No. 0026417. The problem is that there is one case of isolation of relatively small fractions, namely white blood cells, platelets, and neocytes. For example, it is difficult to That's a certain point.

European Patent No. 0026417 describes a method for separating and isolating blood components. The method for doing so is described below. According to this method, after centrifugation, the first layer is pumped out and collected. This pump delivery is controlled by a pressure cushion ( It is carried out by applying pressure from one side to a flexible barber vessel using a buffer (buffer part). There is. The specification describes how the plasma is transferred to an adjacent receiving container. An explanation has been given regarding this.

The blood component separation and isolation device described in the above-mentioned European Patent No. 0026417 The position is the distance from the axis of the centrifuge as it rotates with the centrifuge during use. one or more containment vessels mounted radially apart; It consists of a centrifuge. Each containment vessel is oriented primarily toward the centrifuge axis. including a processing vessel with an outflow opening, the processing vessel having one (or (two or more) receivers are combined with a container to form a closed system. This centrifuge is equipped with a pump mechanism, which pumps the components after centrifugation. It is sent from the container to the receiving container.

From page 816 onwards in Volume 217 of ``Nature'', there is a description of the ``continuous flow'' method. A method is described that uses the method to eliminate the above-mentioned drawbacks. According to this method, The fractions are separated and isolated during centrifugation. Therefore, contamination after centrifugation will not occur in this case.

However, the above method also has the following disadvantages. i.e. supply and drain Requires pipes and associated passage members and rotating seals, which can be costly. The disadvantage is that it results in a more complicated structure. Another drawback is that the single centrifugation process In this case, only one container can be used, which means that the throughput capacity is low. That is.

"LΩJ1 According to one aspect of the present invention, centrifugal separation is performed on the processing vessel, and fraction components are separated from the centrifugal fraction. Disclosed is a method and apparatus for pumping a separate and co-rotating receiving vessel. .

With this method according to the invention, the problems described above do not occur and in a simple manner. The pumping of the components during centrifugation becomes possible, and for this purpose the centrifuge consisting of After the separation time, the pump is pumped during centrifugation using a pump mechanism that rotates with the centrifuge. the entire liquid system, i.e. the receiver is the container(s); The control vessel and coupling pipe(s) keep the centrifuge in equilibrium. It has the characteristic that it rotates at the same time.

The fact that the pump rotates with the centrifuge eliminates the need for costly pipes and circuits. An unbalance in a centrifuge, which makes transverse seals unnecessary, can be caused by (multiple) on the same plane as the processing vessel with respect to the axis of the first layer after centrifugation, or Or, alternatively, for example, by another, also co-rotating, liquid system, the field of the Avoid this by compensating for the weight of the liquid flowing out of the control container at the location. Can be done.

Centrifuge to achieve and maintain a very clear separation surface between N during pumping If a high rotational speed is required, this method can be used to A vacuum is formed in the coupling pipe between the The problem arises that it is inhibited.

According to another feature of the method of the invention, this problem can be solved by overpressuring the liquid in the system during centrifugation. It can be overcome by applying force.

Such pressures can cause flexible container systems that can only expand to a limited extent. This can occur in one embodiment of the method of the invention, where an excess of liquid is introduced. Ru.

Limiting expansion can be achieved by, for example, placing two containers in a closed containment vessel. It can be realized. In this case, pumping excess liquid into the system will cause the flexible walls of the container to It expands to completely fill the containment vessel, after which the desired overpressure in the system causes the liquid to It occurs with just a slight excess. For example, a container made of plastic may be used. A disposable container could be used, e.g. It is possible to introduce into the system from an extra container that is in fluid pressure communication with the supply pipe. I can do it. The container is filled with sufficient liquid and its capacity is related to the expansion potential of the system. When a vacuum is created in the system, liquid is drawn into the system from this container.

Such excess pressure also. Externally increasing the pressure in a system or part of it This can be caused by Of course, to get the desired effect, use less A part of the system, for example a container, has flexible walls, from which excess It is necessary to be able to apply extra pressure to the system.

The above-mentioned method is particularly suitable for separating blood into its components as well as for separating bone marrow cells. This applies especially when

The above-mentioned components of blood are plasma, red blood cells, platelets, and white blood cells, and their respective ratios are Weight is 1.03.1.10.1.05 and 1.07 g/ml. white blood cells It can also be divided into mononuclear cells and granulocytes. Various ingredients with high purity Demand is high for patients with undesirable immunology as a result of blood transfusions and transplants. To avoid the above reactions, it is advisable to manage patients only with the necessary ingredients. It is. Platelets and white blood cells account for only about 1% of blood, and platelets and mononuclear white blood cells Since blood cells and granulocytes must be isolated from this mixture, considerable How to avoid contamination of components with blood cells or cells of other components to a large extent is difficult to achieve, nevertheless the need for such a method is big.

The advantage of the method according to the invention is that the isolated components have a very high purity. The yield or amount of components that can be extracted from a given amount of source mixture is It is considerably more expensive than in the case of the method of more units can be processed and, as a result, more units can be processed per unit time. It has the advantage of being able to separate large amounts of blood.

When separating and isolating blood components, a layer containing platelets and white blood cells and a layer containing plasma and white blood cells are separated. The dividing interface between the layers containing red blood cells and red blood cells should not be disturbed during pump operation. For example, use an arm length of 26 cm and 5o.

It is considered important to perform the centrifugation at a rotation speed higher than rpm. Approximately 80 In Orpm, there is a problem of vacuum generation in the coupling pipe, which increases the pressure in the system. It was necessary to At 2000 rpm, the required overpressure is 6 atm. It was thought that This excess pressure forces more fluid into the system, such as saline. This was achieved by introducing

The invention also relates to a device for carrying out the method according to the invention. This device is , in the radial direction relative to the centrifugal axis Jl1 that rotates with the centrifuge during use. consisting of a centrifuge with one or more containment vessels at a distance of The storage container functions as a storage container or is equipped with a storage container. The container has an outlet opening, the opening being at least primarily radially oriented; A closed fluid system with receiving vessels provided by tubes that are also primarily oriented in the radial direction. and the device is coupled to the container(s) for receiving liquid from the treatment container. It is equipped with a co-rotating pump mechanism for pumping.

Each containment vessel is provided at the end of an arm extending radially from the axis of rotation, for example. It is. A tube extends from each container, and inside this tube is a pump that rotates together with the tube. The pump mechanism causes a flow of liquid to be transferred to the receiving container.

In order for the centrifuge to remain in equilibrium during pump operation, the arm of the centrifugal force am For this purpose, the mass of the end of the pump must always be the same. During the feeding operation, liquid is continuously introduced into the container.

In a preferred embodiment, the pumping mechanism provided in the device comprises a first liquid system (1) a second flexible receiving vessel that fills the containment vessel along with a flexible storage vessel; It consists of a co-rotating liquid system (11), and the device is located outside the containment vessel. The liquid to be centrifuged must be Contains a liquid with a certain degree of The latter, heavier liquid forms a closed system of vessels. The receiver is press-fitted into the containment vessel by the centrifugal force within the vessel (11). When this happens, an equal volume of fluid is forced to flow out of the first system's processing vessel (I) under pressure. . Therefore, the total mass at the end of the arm remains approximately constant.

Here, the two systems indicated by ■ and 11 are the processing vessel (1) and the receiving vessel (■ are located in one containment vessel, and the storage vessel (II) and the receiving vessel (I) are located in the other. are coupled together in such a manner that they are located within a containment vessel.

In principle, a co-rotating pump can It can be placed anywhere inside the aircraft. Therefore, it is possible to move in the radial direction. Therefore, the specific gravity of the conical "cap" placed on the heating container should be 1 minute. If the specific gravity of the two components is between, it can act as a "pump". this If the receiver is located so that the container faces radially inward of the cap, the ingredients are as it flows out from the container into the receiving container.

The cap is pressed radially outward, thereby acting as a pump. .

In the apparatus according to the invention, the processing container and the corresponding receiving container(s) are The problem of equilibrium can be simply solved by housing it in a containment vessel. To avoid creating a vacuum in the system when performing centrifugation at high rotational speeds. The two devices according to the invention both have flexible walls and are located within a containment vessel. containment vessel(s) are used, and the containment vessel(s) When in use, both containers fill the containment vessel and This allows the extra container to be filled with liquid during use to be placed between the containment vessel and the centrifuge shaft. and is connected in fluid pressure communication with the system by an open joint.

Preferably, the additive is a saline solution having a density greater than the various components of the mixed fluid. The replenishment fluid in the replenisher is heated to displace the separated components that are lighter than it. It can also be allowed to flow into the container. Pressure is applied to the container by contacting the container. into a separate pressure vessel or balloon in which the separated components are transferred. can be adapted to suit your needs.

Various designs can be selected for the pump mechanism. in one preferred design According to the invention, the device comprises a screw pump of the usual type rotating together with a centrifuge, which The drive shaft of the pump is equipped with a clutch, for example, which can separate the coupling during centrifugation operation. Located in a more mechanically coupled extension of the rotation axis of the centrifugal process. centrifuge The connection to the shaft of the pump takes place, for example, via the pump housing. Clutch: 1 example For example, by using a pressure plate attached to a fixed point, such as the lid of a centrifugal force machine, It can be separated by separating the drive shaft. Then, the pump housing The pump rotates about its fixed drive shaft, and the pump performs its pumping action.

Other designs of the pump mechanism are also possible. For example, the pump mechanism can be combined with a centrifuge. installed on or near a rotating shaft to separate the heaviest components of the mixture to be separated. It can be made into a cylindrical body (CI+) filled with a liquid having a higher density. This cylindrical body is connected to the flexible receiving container (1) via a pipe. Ru. The receiver is a container (11) with a closure provided at the end of the arm of the centrifugal force 1l11 machine. together with a flexible source container (1) of the mixture. centrifugation During this time, this liquid flows to a receiving vessel (1) located within this containment vessel. This capacity Once the container is filled, the container (I) is compressed and the liquid inside it is pushed onto the centrifuge shaft or or into a nearby receiving container (1). Centrifuge loses equilibrium In order to avoid It is necessary to select a liquid with a density slightly greater than that of the liquid.

According to some aspects of the invention, to achieve accurate component isolation of fluids being processed, For this purpose, various mechanical elements and Both forms are adopted.

The method according to the invention requires, firstly, that as much as possible of each component be produced in a high purity per amount of disintegration liquid. So that you can get a large amount.

To be able to "process" as much blood as possible in a single centrifugation process and second, to accomplish one centrifugation process in the shortest possible time. They argue about things. In order to accomplish W41's argument, it is necessary to It is important to make optimal use of the space available inside the Centrifuge II machine for the layout of the be.

One method according to the invention provides a pump machine that does not occupy space in the sense mentioned above. We start from the recognition that it is possible to achieve the desired structure. This method is used for this purpose in.

Reduces the volume of the bar by pushing the radially outward walls of the bar inward. It has the characteristic of realizing pumping by doing so. This feature has two functions. The first means for realizing this feature according to the present invention can be achieved by Under the influence of centrifugal force, the outer wall of the barber is a ridge formed at the bottom of the containment vessel. This is carried out by moving the processing container radially outward so that it is in contact with the In a second means of achieving the above characteristics, inward repression is Move the ridges on the bottom of the container radially inward so that they fit inside the outer wall of the container. It is carried out by pressing in the direction.

Furthermore, in order to shorten the period of one centrifugation process as much as possible, it is necessary to It is important to always do this at the highest possible speed. Several layers are relatively thin Therefore, there is an upper limit to the speed at which the few layers can be pumped. child When the upper limit of is exceeded, the number of layers is .

When it reaches the outflow opening, it "breaks" and the red blood cells are pumped out along with the soft calendar. It will end up being lost.

In order to achieve an optimal pumping process with respect to duration, the present invention According to another feature of the method according to Set relatively high when pumping large volumes of components, and set relatively high when pumping large volumes of components, and when pumping relatively small volumes. When pumping the product component, the speed is set relatively low.

In the apparatus for carrying out the method of the present invention, the side wall of the processing vessel is approximately funnel-shaped and has an outflow opening. converging towards the mouth and further from the radially outer side of the container at an adjustable speed. It has the feature that it is equipped with a mechanism for reducing the volume of.

Funnel shape at the radially inner end allows for effective separation of components Become. If only a small amount of one calendar remains in the container, the funnel shape will The layer thickens as it approaches the outlet and pumps without mixing with the next layer. Can be sent. This is especially true when the overall thickness is very small. This applies to several layers that have not been used.

The mechanism for adjusting the speed of compressing the container is effective in obtaining the optimum speed. . That is, the amount of flow from the container can be adjusted for each layer depending on the thickness of each layer.

The funnel shape is formed from a rigid material at the radially inner end of the flexible container. In a preferred design of the device according to the invention, there is no leakage at the inner end of the container. A rigid, bowl-shaped cap is fitted. This cap has an opening and The skirt of the lever is oriented from the outer end to the inner end and is at least equal to the diameter of the inflow vibrator. open across the width. The opening formed in the skirt allows the cap to be Can be installed in a container with a permanently attached outflow vibrator. like this Preferably, the cap is conical.

The design features of the device according to the invention are suitable for methods in which the processing container moves radially outwards. According to the indications, the outer wall of the containment vessel has a raised part inside, and this raised part is In the parked position, the inside of a primarily cartridge-shaped housing containing the side walls of the container The containment vessel is in contact with the radial outer wall of the containment vessel installed in the It is movable by sliding radially along the inner surface of the side wall of. take the container With a rollable cartridge, the container can be moved within the containment container. Housing average Preferably, the caps and caps are connected to each other by threaded closure means.

The space that the centrifuge must provide for the containers of blood to be separated should be According to a further advantageous design of the device, the walls of the housing are externally radially inwardly from the convergence of the housing, while the containment side walls also converge inwardly in a substantially parallel manner, and the wall or portion of the wall of the containment vessel is It can be removed over a width corresponding to the radius or diameter. This kind of containment vessel , can be arranged as sectors of a flat disk around the axis of the centrifuge. can.

In order to create a space for the container, the container for the storage container is formed into a conical shape. It is preferable to Can be installed on the outside wall of

In a structure with a convergent-shaped housing within a convergent-shaped containment vessel, the housing When moving the housing in the radial direction, the housing must be guided. In devices of such design, one end is fixed to the inward facing part of the containment vessel and the other end is portion of the inner end of the housing as well as the outer periphery of the containment vessel extending approximately radially inwardly. radially oriented slidably coupled to a guide surface on the inner dorsal surface A support 44 @ J/q is constructed from a rod that has been In this case, two The talophyte between the edges corresponds to the distance between the housing walls on the outer part of the housing and is less Extends inwardly at least to a height beyond the ridge.

It also controls the rate at which the fluid is pumped and therefore the rate at which the outer wall of the chamber is compressed. According to one design of the mechanism used for this purpose, its characteristics As a characteristic, the outer surface of the container extends partially in the form of a ring over the bounding surface of the ridge; Then, below this part and around the ridge, there are Two or more inflatable rings are installed and independently adjustable A valve having a flow opening is provided. This design allows two pump speeds becomes. In the first phase, when pumping plasma, the speed can be high. Wear. The valve of the first air-filled ring is in a state where the other valves are closed. can be adjusted to increase the flow rate. When several layers reach the outflow opening At this point the speed must be reduced.

The previously open valve must close, and then the other valve with a smaller opening opens. It will be destroyed. This results in lower pump delivery.

Opening and closing of the valve can be electronically controlled using a sensor, for example.

Instead of using centrifugal force to move the barber vessel radially outward, An object can be used that presses the outer surface of the vessel inward, in which case the outer wall For example, by providing an opening in the A ridge having a matching inner surface can be moved radially inward.

In particular, components that together form a relatively small soft-layer fraction are separated, isolated, and A problem arises when #I is in state. The components of these layers include white blood cells and blood cells. Contains platelets. This also means that the thin top or lightest layer of the red sphere, immediately This also applies to the calendar of Chi Neo Site (Shin Akadomari Ball). The receiver has good separation in the container. In order to achieve this and to enable isolation without contamination after separation, Uke has special requirements for containers.

Such a requirement in the design of the device according to the invention.

The receptacle consists of a tube connected to the outflow opening and open at the outer end indoors, and In this case, the volume of the tube is determined by the platelets, white blood cells, and possibly white blood cells present in the blood or bone marrow. should be at least equal to the volume of the neosite, while the diameter is generally At rest, the tubes are selected to be large enough that no mixing of the components occurs, and during centrifugation, the tubes are barber means for positioning the tube in such a way that the g-value of the tube decreases along the length of the tube. Installed on the radially inward part of the container.

In particular, the tube has a centrifugal force gradient sufficient to maintain separation between the different fractionated blood components. It is desirable to support the tube in such a configuration that it occurs along the tube.

In the chamber, plasma is first collected. Thereafter, the radially inward end of the source vessel Several layers are pumped into the tube, divided into two components. These few layers Contains two components, and in some cases neocytes or new red blood cells. may be included. These two or three are positioned like this. Note that 1 minute is the To ensure that the centrifugal force inside the tube remains intact.

It is necessary that the flow rate decreases continuously in the direction of inflow into the tube. this thing In practice, when the tube is stored wound in some way, for example on a reel, , this storage must be carried out in a manner that serves the above purpose. in this case is located radially inward from the outflow opening and preferably the source Attach onto a rigid cap on the inner end of the container.

In order to easily collect the components to fill the tubes sequentially, place the tubes where the specific components are located. Advantageously, it is connected to a closable auxiliary chamber provided in the area of the tube. This attached room By opening the inlet to the chamber, the relevant components can flow into this chamber, The room will then be closed again.

One object of the present invention is to The purpose is to make it possible to isolate even the most important parts in an effective manner. Books suitable for this purpose The device according to the invention is characterized in that the outflow vibrator collects at least the leukocytes, platelets, present in the source container. and optionally flexible with a volume equal to the total volume of the neocyte (new red blood cells) During centrifugation, the centrifugal force within the tube is applied to the centrifuge arm or arm. To store the tubes in such a way that they decrease continuously from the outside to the inside when viewed in the direction of the beam. It is characterized by being equipped with the following facilities.

The tube forms a connection between the source vessel and the receiving vessel(s). Inside the heating container After # minutes of the components, the liquid is centrifuged into the source vessel with the axis of the molecule 6 machine Relatively light blood when pumping in one direction or the other through the orifice. Serum first enters the tube and from there flows into the receiving vessel. After that, several layers Platelets are on the inside, white blood cells are on the outside, and neocytes are on the outside. or at least flow into the tube in such a way that neocyte-rich red blood cells are located.

During centrifugation, to ensure that the centrifugal force in the tube decreases continuously in the flow direction In one design example of the device according to the invention, the chain tube is perpendicular to the axis of the centrifuge. It is wound on a reel that has a straight axis and a circular cross section perpendicular to the axis. In the case where adjacent turns of the tube continuously form positive angles with the The size of the angle also depends on the tube thickness, reel diameter, and distance to the centrifuge axis. Make it change depending on the distance.

Here, "iso-g line" refers to a closed line that surrounds the surface of the reel and has an equal centrifugal force. These lines are not the same as the circular cross section of the reel. stomach.

In addition, the equi2 surface is the surface of a cylindrical body whose central axis is the axis of the centrifuge. The zero magnitude glji is defined as .

Represents the intersection between the surface of the reel and two surfaces such as those mentioned above.

Therefore, the centrifuge # is the curve surrounding the reel of equal radial distance from the axis of the machine. Ru. Therefore, the iso-g line on the reel roughly outlines the saddle shape centered on the reel. When winding a tube on a spool-like reel so that it always forms an angle with the zero isog line In this case, a centrifugal force gradient acts on the contents of the tube to remove the components that would otherwise be present. Mixing is prevented.

In one embodiment, the turns or turns of the tube, viewed in the axial direction of the reel, include: The increase in the thickness of the edge flange over l turns in the winding direction is equal to the thickness of the tube. Therefore, they are located adjacent to each other.

In order to achieve good functioning of the device according to the invention, it is necessary to pump The dimensions given to the tube in which the components to be isolated are located are very important. .

The device according to the invention is used for the separation and isolation of leukocytes and platelets from blood. In the preferred design of such equipment in cases where the volume of the tube is the internal diameter of the tube is at most 5 m. is chosen to be equal to m.

If the diameter of the tube is larger than 5 mm, after the centrifugation process is completed, It is considered that relatively large contamination occurs on the interface surfaces of the two separated components due to mixing. It will be done.

The device according to the invention is capable of separating and isolating leukocytes and platelets from blood. If the tube is also used for the separation and isolation of neocytes (new red blood cells), The volume must be at least 11% of the volume of blood in the warm container. The diameter of the tube must be a maximum of 5 mm to avoid contamination on the surface This requirement does not apply to the length of the tube where neosite is present, but rather For example, if the volume of blood in The first member may have a diameter of . Divide the tube into two parts like this. The advantage of dividing the material into pieces is that it can be made from pipes that are as long as 250 cm and difficult to handle. The point is that there is no need to do any work.

If the source material is bone marrow rather than blood, the volume of the tube will increase after collecting several layers of components. Must be at least 4% of the bone marrow outflow volume in order to separate There was found. The reason is that bone marrow contains a relatively large number of white blood cells. . Less stringent requirements are imposed on the diameter of the canal; in the case of bone marrow, the diameter of the canal is The diameter can thus be, for example, 1 cm.

Instead of a thin-walled tube with reels, it contains platelets, spherules and neocytes. A tube can be used that has an overhang at the point where the particles converge. These overhangs are The tube is divided by a narrowing, and can be closed and isolated by tightening at this point.

Relatively small volumes of components may contaminate each other or platelets may become contaminated with plasma. Alternatively, neocyte-rich areas may be contaminated by neocyte-poor blood. It is important to maximize the rate at which these components flow into the pipe in order to reduce It is essential. In any case, the separation and isolation process can be completed within the shortest possible time. According to another feature of the closed system according to the present invention, in order to Controlling the flow rate or proportion of components flowing out of the container according to the components to be discharged It is advantageous to have means capable of or to operate in cooperation with such means. It is. Such measures may depend on the components whose passage is detected, e.g. Sensors may be provided to adjust the amount or rate.

This book is useful when you need to separate neocytes, white blood cells, and platelets. In a preferred device design according to the invention, the treatment container has a specific gravity smaller than the neosite. the spheroid preferably has a specific gravity greater than that of white blood cells, and When the above-mentioned spherical body comes into contact with the outflow opening from the control container to the pipe, the spherical body opens the outflow opening. is given a shape such that it is partially closed.

When several-layer components and subsequent neosites flow into the tube, small spherical bodies partially closes the outflow opening, so that the subsequent influx of red blood cells into the tube is slow. This reduces the chance of mixing within the pipe.

In another preferred design, the means comprises a first blood component present in the device. A sensor that records the outflow of the ball and cooperates with the device to adjust the outflow amount.

During centrifugation, the flow rate is also reduced when using a chamber with the shape of can be done. At the end connected to the outflow vibrator, such as a cleaning container, to reduce the flow rate when platelets begin to flow into the flow vibrator during centrifugation. A barber vessel having a narrow portion is considered. After plasma flows out, it remains in the laboratory container. When a collection of red blood cells, white blood cells, and platelets is forced to change its shape, It has such a viscosity that the flow rate decreases with the amount of energy supplied.

By adjusting the shape that forms the narrow part to this viscosity and the amount of blood cells as described above. This achieves the effect that the outflow volume begins to decrease as soon as platelets reach the outflow opening. can do. A narrow area is defined as a narrow area where a collection of related ingredients is located within the relevant part of the container. Any shaped part of the vessel that results in increased flow resistance when flowing into the For example, the outlet side of a cylindrical container can be conical. corner of the wall The degree is selected such that the desired reduction in flow rate is achieved.

Various methods can be applied to isolate the components present in the tube at any given time. For example, pipes can be separated using clamps that tighten at interface surfaces between components. The contents of each section are then placed under pressure in a chamber provided for this purpose. It can be made to flow in.

In a preferred design according to the invention, the section of the tube in which the neosite is collected is It is connected in a closable manner to an auxiliary chamber in which, after stopping the centrifugation process, Neosites are collected. There are also rooms like this. For platelet chamber and white blood cells It can also be used in conjunction with a ward room.

In order to isolate the different components present in the tube, a connected auxiliary chamber (single or or plates) to which the filled tube can be fixed after unrolling. It is advantageous to use a stand. Such a board has a A clamp is provided that is slidable on a rail extending over the rail. These clubs The pump is clamped onto the tube at the point in the tube where the dividing plane between the two components is located. This allows the above-mentioned divisions to be formed.

In the following, embodiments of the device according to the invention will be schematically illustrated, as well as those occurring during the implementation of the invention. The invention will now be explained in more detail with reference to the drawings which illustrate the situation.

Brief description of the drawing FIG. 1 is a horizontal cross-sectional top view of an apparatus illustrating one aspect of the invention.

The second is a side view of the device in FIG. Figure 3 shows the storage volume after centrifugation for a period of time but before the start of pumping. A diagram showing details of the inside of the vessel.

FIG. 4 is a diagram showing the situation during pumping operation.

Figure 5 is a cross-section of a centrifuge schematically showing two different designs of centrifugal units. View from above perpendicular to the axis of the centrifuge.

FIG. 6 is a detailed cross-sectional view of one of the centrifugal units shown in FIG. 5; Figure 7 shows an alternative to the one shown in Figure 5, which has a raised pump mechanism. Similar detailed cross-section of the centrifuge unit, Figure 8 shows the centrifuge in the center of the conical centrifuge unit shown in Figure 7. A diagram showing a cross section parallel to the axis.

FIG. 9 is a diagram schematically illustrating a closed system of source and receiver; Figure 10 shows the assembly with a cap placed on the radially inward end of the This is a reel that forms the body, and the receiver is a reel for winding up the tube part of the container. The diagram shown in the figure.

Figure 11 shows that the tube consists of two parts of different thickness and that the container contains a sphere. a diagram schematically illustrating a closed system according to the invention, and Figure 12 shows a tube section with a closable auxiliary chamber, and the entire tube is clamped. Schematic representation of a closed system according to the invention fixed on plywood with rails provided This is a diagram.

1] J In Fig. 1, the containment vessel l is equipped with a storage vessel 2 and a receiving vessel 3. Both containers 2 and 3.

They are interconnected by pipes 4 and made of plastic, for example disposable. It is made of flexible material such as. The pipe 4 is located above the centrifuge shaft 5. It passes through a pump 6 attached to. In Figure 1, there are four "arms" A centrifugal separator equipped with

Note that the pipes 4', 4'' and 4'' shown in the figure are from the centrifuge arm shown in the figure 1. It corresponds to the outer arm or arm.

The pipe 4 is provided with a valve or closure 7, which allows the constituents to The pipe can be plugged when separated from the mixture and passed through. The closing part 7 is For example, a light source and a photovoltaic device placed around the pipe to detect the passage of a dividing layer between the components. responds to signals from an “eye or sensor” that may consist of a detector and .

Connected to the pipe 4 is an extra container 8 filled with physiological saline.

This solution is sucked into the system when a vacuum begins to be generated in the system at a rotational speed of . The control vessel 2 and the receiving vessel 3 partially absorb the excess liquid fed into the system from the vessel 8. When the body "fills" the containment vessel as a result, there is an undesirable vacuum offset within the system. Such pressure is generated.

, FIG. 2 shows the same device in side view, the reference numeral 9 being at the end, centered on the axis lO. It also represents a centrifuge arm with a containment vessel movably attached to the The reference numeral 11 represents the drive shaft of the pump, and this drive shaft is connected to the pump housing 6. It is also mechanically connected to the shaft 5 of the centrifuge. This drive shaft 11 is a pressurizing part material 12 can be used to separate during centrifugation.

Figure 3 shows the situation inside the containment vessel 1 after centrifuging the blood for a certain period of time. The fluid is red blood cells13. Plasma 15 and blood in the 1j14 form between them It is separated into so-called "several layers" consisting of platelets and white blood cells.

Figure 4 shows the situation after the centrifuge pumps out for a certain amount of time. 15 is a zero-order component that flows out of the processing container 1 and flows into the receiving container 3 as the first component. When several layers are located in the narrow pipe 4 and form a relatively thick layer. By stopping the pump operation, the platelets 14'' and the remaining white blood cells 14' are separated. It becomes possible to perform separation between

In Figure 5, the reference numeral 16 represents the centrifuge shaft; Units 17 and 18 are installed. One centrifuge generally has one Centrifuge units of type 1, e.g. all units of either type I or type 2. It is equipped with Here, for convenience of explanation, "type l" corresponds to the design of unit 18. Accordingly, "type 2" shall correspond to the design of unit 17. centrifuge unit 17 and 18 each have a radially outer wall 21 or 22. It is composed of a container 19 or 20. There is nothing inside the containment vessel 19 or 20. Each has a flexible structure for the blood to be centrifuged and is made of plastic, for example. There is a processing container 23 or 24 that is The walls of containment vessels 23 and 24 are , facing the outflow opening 25 or 26 and converging at the radially inner end in a funnel shape.

The outflow opening 25 or 26 is connected to a receiving vessel 27 or 28, part of which is shown. open inward. To ensure the sterility of the contents, and the receiving container are connected to each other via an outflow pipe. Reference numbers 29 and 30 are the axis of the centrifuge relative to the outer walls 31 and 32, respectively, during the pumping operation. The ridges 29 of the units of type 1 represent ridges moving in the direction of , i.e. inwardly. , permanently connected to Jj121 outside the containment vessel. During pump delivery operation, A container 28 located within a housing that moves with a container (not shown) moves outwardly. and located on each side of the ridge 29. In the case of type 2 units, the raised portion 30 is provided outside the outer wall 22 of the containment vessel 20.

This outer wall 22 is provided with a hole through which the raised portion 30 is centrifuged. Move in the direction of machine axis 16.

Thereby, the outer end of the container 24 can be pushed in.

It should be noted that means for moving the raised portion 20 inwardly is not shown; this movement can be done, for example, by Various methods are possible for pumping, which could be done using fluid pressure. For example, by pressing the ridge into the outer edge of the container or by pressing the container against the ridge. Pumping can be performed by pressing, and any of the types of containment vessels mentioned above It can also be applied to

When using a type 1 centrifuge unit, there are two types of centrifugal separation units: For example, type l, which allows better utilization of the space inside the centrifuge In the case of units, 12 standard units can be placed on one disk. I can do it.

Figure 6 shows details of the design of the type 2 centrifuge unit in the same cross-section as in Figure 5. be. As shown in Figure 1, the processing container 23 has a wall portion 33 that converges in the shape of a funnel at the inner end. do. This wall section is held in this shape by a cap 34 that rests on it.

A hole 35 is provided at the inner end of the cap 34, and the hole 35 is designed to prevent the flow from flowing when in use. located above the outlet opening 25 so that the outlet pipe 36 can pass through the opening. It has become. To enable the cap 34 to be placed on the processing container 23. For this purpose, the jacket or skirt of the cap extends outwardly from the hole 35. It shall be provided with an opening (not shown) that allows the

The processing vessel 23 is supported by a housing 38 on its side walls 37, preferably over its entire height. has been done. The housing 38 extends from inside to outside along the inner wall 36 of the containment vessel 19. In the example shown, it is possible to slide in the direction of 0.

Cap 34 and housing 38 are connected to each other by a threaded closure 40. There is.

The receiver is composed of a pipe 41, and one end of the chain pipe 41 is connected to the outlet opening 25 of the container 23. and the tube 41 is connected to a spool mounted on the cap 34. Alternatively, it is wound on the reel 42. The other end of chain tube 41 has one or more ports for components. or multiple collection chambers (not shown). The container contains the following ingredients: (passing), a sensor 43 and a valve or closing part 44 are provided to detect this. It is being

In order to be able to divide the blood in the treatment container 23 into its components, the blood is first , 0 separation in which the closure part 44 is closed and blood is centrifuged without being pumped out of the container. When this is completed, the closure 44 is opened, and the centrifugal force causes the container 23 to open in the containment vessel 19. It moves outwardly by sliding along the inner wall 39 with its housing 38. container deflection The flexible outer wall then abuts the ridge 29 and at points 45 and 46 the ridge extends in the form of a ring on the sides of the container 23, whereby fluid is forced out of the container 23. this In this way, after the first component, i.e. plasma, has been pumped out, the next This is followed by several layers, which are under pressure inward during plasma pumping. These few layers will eventually be located at the innermost tip of the funnel. Leakage Due to the bowl shape, the surface area of these layers decreases and therefore their thickness increases. but However, during pumping, to avoid the fluid of the next component flowing out together. The pump delivery rate must be relatively low, i.e., the pump delivery rate must be relatively low, i.e. It needs to be lower than the required speed or amount.

Valves 49 and 50 are provided to allow adjustment of the above-mentioned outflow rate or amount. Two inflatable rings 47 and 48 surround the ridge 29 and extend forward and backward. It is located. The constrictions of rings 47 and 48 and valves 49 and 50 allow blood plasma to As long as it is being pumped, a relatively high rate of contraction will occur from ring 47. However, when it is the turn of several layers, the ring 48 is dimensioned so that it contracts at a slow speed. can do.

Figure 7 shows the cross section of the centrifugal separation unit 17 shown in No. 512I in more detail. Note that the difference from FIG. 5 is that the relative movement of the outer W 32 with respect to the raised portion 29 is different from that in FIG. 7. In this case, as shown in FIG. Instead of being pressed against the protruding portion 29, the processing container 24 is pressed outward. This is the point that can be achieved. As already mentioned, the advantages of the container shape shown in Figure 7 are: The filling degree of the centrifuge is improved.

However, in order to enable pumping even with centrifugal force, therefore, Some special means are required to allow the container to move outwards. be done. It is equipped with converging walls 20 to make use of the available space. In the containment vessel, there is a convergent housing 53 supported laterally by a convergent housing 53. A container 24 is arranged with a wall 52 that has walls 52 . In the starting position the housing is , the wall thereof is in a position where it abuts the wall of the containment vessel 20. This housing 53 It is necessary to guide the housing 53 when it begins to move under the influence of mental forces.

For this purpose, a guide 54 is attached to the containment vessel 20, which guide 54 also . such that the wall of the housing 53 has a cartridge shape that slides along it. Preferably, the housing further has a forward end retracted as indicated by reference numeral 56. is fixed to the container 20 and the other end is fixed to the inner wall 57 of the housing 53 in a similar manner. radially guided by fixed rods 55. Note that the rod 55 is connected to the housing. It protrudes through the inner wall 57 of.

Reference numeral 58 indicates an inflatable ring corresponding to rings 47 and 48 shown in FIG. This diagram schematically shows the

Elements 43 and 44 respectively accommodate the previously described sensor and closure valve in the container. The tube 41 of this receiving container is wound on a reel (not shown) as shown. ing.

No. 713 shows a state in which the housing is located at the outermost position.

8 [2I is relative to the centrifugal axis of the centrifuge unit as shown in Figure 7. Note that parallel vertical sections are shown.

This centrifugal separation unit has a conical shape. The advantage of this conical shape is that centrifugation Viewed in the direction of the axis 85, the collection chamber of the receiving vessel can be located above the containment vessel; This ensures that the maximum amount of space is available for the container containing the blood to be centrifuged. There is a certain point. Collection chamber 59 for Jf11 plasma as well as a second chamber for platelets A collection chamber 60 is shown in the figure.

Next, referring to Figure 9, Uke explains the separation of plasma and platelets in the chamber. do. In addition, FIG. 9 shows the tube 41, the plasma collection chamber 59, and the platelet separation auxiliary chamber 6. 2 is a diagram schematically illustrating a closed system of a processing container 24 and a receiving container composed of 0. blood When liquid is present in container 24, this container is closed with snap connection 68. The system is then centrifuged. The snap connection 68 is then destroyed. , in which the closing part 44 (see Figure 8) takes over the closing function instead of the snap connection. After several layers have been formed, the closure 44 is opened, where the plasma and several layers can be isolated as described above. When the platelets pass through the sensor 43 Then, the closing part 44 closes again. This means that for the last centrifuge unit Once done, stop the centrifugation and remove the system from the centrifuge. The tube 41 is then unwound 65. The tube 41 is then clamped 65. to the boundary of the different components. By placing 66, 67 and 68.

Divide into sections corresponding to locations where different components exist. Close the auxiliary chamber 60 from the pipe 41 By opening the snap connection 61, the component 621, e.g. 60 under pressure.

Platelets, white blood cells and neocytes remain separated from each other within tube 41 In order to ensure that the g value acting on the fluid during centrifugation is It is necessary to decrease continuously and evenly. For this purpose, the tube has an outflow opening should be stored in a special manner, radially inward from the ground. tube or adjacent turns. or a reel that is wound to form a turn has a special shape for this purpose. Must.

FIG. 1O shows that in order to comply with the above requirements regarding the g value, What kind of reel is reel 42, which is shown in the position where it is permanently coupled to A perspective view shows what should be done. The tube 41 has a protruding edge from the cap 34. are regularly wound around a reel portion 69 located between 55a and 55b. .

FIG. 10 also shows that the jacket or skirt portion 71 of the cap 34 has a The open lower 2 allows the cap 34 to be arranged on the processing container 24. It shows you how to do it. To achieve correct winding, the reel must be The thickness 74 of the tube 41 increases continuously in the winding direction from the point 73 where the tube 41 enters the reel. large, so that the reel increases by the diameter of the tube for a thickness of 75 after one winding. is configured to add

As shown in FIG. 10, the reel 42 has a reel axis 80 and a conical It is connected to the mold cap 34. During use, the cap 34 should face inward of the processing container. fall onto the surface. flexible tubing (not shown) connected to the outflow opening of the processing vessel; is wound in adjacent windings or turns around the reel surface 69; The winding is limited by projecting edges 55a and 55b.

The tube enters the reel via the open lower part 3. ! The thickness 74 of the i portion 55a is the same as that of the open lower 3. The thickness increases continuously from 75 to 75 along the winding direction 81, in which case the thicknesses 74 and 75 The difference between 0 and 0 corresponds to the diameter of the tube used when performing winding on a wound tube. , the chain pipe abuts against the edge 55a, and the following winding or turn always follows the preceding turn. The helical shape of the tube can be seen in a simple manner on the reel surface 69. A winding is achieved, whereby the g-value inside the tube decreases continuously. to reel axis 80 The shape of the reel surface 69, which has a circular cross section in a direction perpendicular to the The distance of the tube from the beginning of the tube to the centrifuge axis located perpendicular to axis 80 It is determined by the condition that it decreases continuously until the end.

In FIG. 11, a closed system according to the invention is used for blood as source material. This is abbreviated as the case may be. Before centrifugation, the blood to be separated is placed in Exists within 6. After the desired separation is carried out in this container by centrifugation, the The contents of the container are pumped out of the processing container via the outlet opening 87. First, blood Serum flows out through flexible tube 88 into receiving container 89 . The tube 88 has a maximum The inner diameter is 5 mm and the content is preferably at least 1% of the total volume of the source material. a first portion 88' having an inner diameter of, for example, 1 cm and whose contents are preferably small; 88'' of W42, which is about 10% of the total volume of the source material.

Several layers are collected in tube section 88' and neosite is collected in tube section 88H. Ru.

The exact volume of the tube sections 88', 88'' is 1 section for a standard source blood bag 86. 88', 88°° is platelet (or neocycline) present in normal human blood. g) selected to retain all fractions.

If we refer to the spatial position radially outward from the centrifuge axis as (upstream), then As can be seen, the larger diameter tube section 88'' is placed upstream of the smaller diameter section 88' during centrifugation operations. It will be supported by the side.

Reference number 104 is smaller than the specific gravity of neocytes, but larger than the specific gravity of white blood cells. This spherule 104, which represents a spherule with a high specific gravity, is the most concentrated during pumping. partially closing the outflow opening 87 when the subsequent leukocytes pass through the outflow opening 87; The runoff velocity is reduced and, as a result, contamination of the surface of the connected soft layer component is reduced. is prevented.

supported to provide a centrifugal gradient to effect or maintain separation of fraction components. 1 separation after removing one or more tube sections from the centrifuge. Final isolation of the material is facilitated by rolling out the chain tube onto a flat table or plate. will be implemented. Visually identify and identify boundaries between adjacent fractions or components in the tube. Each fraction is isolated by placing pinch clamps at appropriate locations.

The isolated fractions or components are then preferably placed in separate chambers. A suitable configuration for this purpose is shown schematically at 1212I.

At UA 12, the closed system shown in FIG. 11 is again connected to the source vessel 86. flexible tube 8 8 as well as a receiving container 89. In this design, the flexible tube 88 has one uniform diameter. After exiting the pump for a period of time, the components, i.e. blood The platelets, leukocytes and neocytes are present within the tube, separated from each other. child Trowel isolation must be carried out; this isolation involves clamping the tube 91.921, Divide into sections 95, 96 and 97 corresponding to different components using 93 and 94. This can be achieved by The content of each component is 1 then 1, for example, auxiliary chamber 98.9 Collected at 9. Of these auxiliary rooms, only two are shown in the figure. Applicable The auxiliary chamber has a snap connection 100. It is connected to related areas through lots.

In order to make it possible to easily isolate the components that are present in the tube and separated from each other, In order to In the unrolled position on a plate 102 with rails 103 that can be moved Can be fixed.

These clamps are then attached to the tube at the dividing surface between the components located within tube 88. Tighten, thereby forming the sections 95, 96 and 97. After that The contents of the compartment can be forced into the auxiliary chambers 98 and 99.

If a tube with an overhang is used instead of a thin-walled tube with reels, The sections 95, 96 and 97 above correspond to these overhangs. Overhang The diameter of platelets is 2 cm (inner diameter 1.5 cm), and the diameter of white blood cells is 3 cm. m (inner diameter 1.5 cm), and in the case of Neosite, 4 cm (inner diameter 4 cm) ), the plate or platform on which the clamp is moved must in this case It's not even needed. This is because the tightening is applied to the part of the pipe located between the overhangs. (1 am length).

In either case, source containers, tubing and overhangs or auxiliary chambers must be kept in a closed, sterile It was formed as a system of Tubes and chambers without are closed off from the actual components. That is, for example, a snap connection At the continuation, the tube 88 connects to a source container 86 into which blood is initially removed from the donor. When centrifuged for separation purposes, the snap connection breaks. The separated plasma can flow through the tube 8B into the plasma receiving chamber W89. 1 Separated white blood cells, platelets and possibly neocytes are collected along the length of tube 88. If closed, destroy the other snap connections that isolate the auxiliary chamber 98.99. Then, the separated components are injected into these auxiliary chambers, and then these auxiliary chambers are It can then be closed, for example by heat sealing.

In this way, blood withdrawal, separation and isolation of blood components can be carried out in a closed sterile takes place within the environment of the state. This results in one separation of soft! The lifespan of components is longer than that of conventional composites. It is longer compared to the isolation method that goes through a number of steps.

The invention has been described above in connection with specific embodiments.

This description is for illustrative purposes only and the invention is not limited thereto. It will be understood that, from the above description, various modifications will occur to those skilled in the art. However, such modifications are within the scope of the present invention.

of Special edition Showa 63-501482 (15) fig, 7 fig, 8 international search report lmeunlmAal　Aell(ll,.,.PCT/NL　861000 02ANNEX To THE I) TTERIJATrONAL 5EARC HRE? ORT 0NUS-A-440507920109/83 NonaU S-A-453193230107/85 NonaUS-A-3519201 07107/70 None

Claims (46)

[Claims] 1. one having a volume not exceeding about 10% of a biological mixture such as blood or In a method for separating multiple fractionated components using a centrifuge, The biological mixture in the source container is centrifuged at a rate and time that results in the desired component phase separation. separate the heart, Maintaining pressure within the source container such that components flow out of the source container and Perform the centrifugation and maintain the pressure, and the separated components will be centrifuged along the length. pumping into a conduit that is positioned such that a centrifugal force gradient of Method. 2. The conduit has a volume sufficient to receive the separated components and the pumping stage And furthermore, The flow from the source vessel is controlled to keep the separated components within the conduit and Claim 1 comprising the step of stopping the centrifugation when the component is located within the conduit. Method described. 3. By winding the conduit onto a spool having a radially oriented axis What is claimed is a flexible tube having a length oriented opposite a centrifugal force gradient. The method described in Section 2. 4. The step of bombing the components involves transferring separated layers of the components from the source container into a conduit. A funnel-shaped cap is provided above the outlet of the facial container to convert it into a columnar body. The method according to claim 2 or 3. 5. The conduit is selected to be large enough to accommodate substantially all of the desired fractionated components. volume and diameter small enough to prevent remixing when centrifugation is stopped. The method according to claim 3 or 4. 6. The conduits each have a volume sufficient to receive separate and distinct separated components. A method according to claim 5, comprising a plurality of sequentially arranged portions having . 7. The source container has a flexible outer wall and maintaining a positive fluid pressure against said wall. Claim 1 further comprising the step of applying pressure to the flexible wall by applying pressure to the flexible wall. How to put it on. 8. Pressure is generated by pressing under centrifugal force against the ridge of the source container and a fluid-contractable support adjacent the ridge and in contact with the flexible wall; controlling relative movement of the container and the ridge by controlled contraction of the retaining portion; The method according to claim 7. 9. supporting the source container within an inflatable housing and applying pressure; is achieved by moving the ridge into the housing against the flexible wall of the container. The method according to claim 7. 10. The pressure-maintaining step moves the fluid system, including the source vessel and conduits, into a confined volume. by providing a replenishment fluid container in pressure communication with said fluid system. A method according to claim 1, which is accomplished by: 11. Used in centrifugation to separate fractional components of sample fluid mixtures such as blood a flexible source container having an outlet; at least one receptacle; connected to said outlet and at least one receiving vessel, said source vessel and said receiving vessel; a conduit which together with the container forms a closed fluid system for transporting fluid from said source container; and a non-zero centrifugal force gradient along said conduit to inhibit mixing of fractionated components during said transfer. and support means for supporting the conduit within the centrifuge to maintain alignment. Device. 12. The support means is further configured to direct a layer proximate the outlet of the sample fluid from the source vessel into the conduit. 12. Apparatus according to claim 11, further comprising cap means. 13. 13. Apparatus according to claim 12, wherein the capping means is funnel-shaped. 14. The cap has a slot extending along the length of the module to allow the conduit to pass therethrough. the cap onto the source container without disconnecting the conduit from the outlet. 14. The device of claim 13, wherein the device is adapted to be worn. 15. Claim 13, wherein the support means is detachably attached to the cap means. 15. The device according to item 1 or item 14. 16. Limit the flow rate from the source vessel to control the transfer of components within the conduit. further comprising means responsive to the presence of the desired fractionated component at the outlet of the source vessel to 14. The apparatus according to claim 13, which comprises: 17. The response means includes a float box having a size to partially block the outlet; The target cell should have a specific gravity intermediate between the specific gravity of the desired fractionated component and the adjacent fractionated component. Scope of Claim 16. The apparatus according to item 16. 18. The response means comprises a constriction in the outlet region, the constriction allowing the desired fractionated component to exit. When flowing into the region, the fractionated components are arranged such that the flow rate through the outlet is reduced to the desired flow rate. 17. Apparatus according to claim 16, having a shape adjusted for minute viscosity. 19. The support means may include a spool surface for winding the conduit over the mole. The device according to item 11. 20. The source container has a characteristic volume of one fractionated component, and the conduit substantially covers the characteristic volume. sized to accommodate said components, such that said conduit serves as a receptacle for said components. 20. A device according to claim 19, which performs the function of: 21. flow control means for the source vessel to control the flow rate of the fractionated components within the conduit; 21. The apparatus of claim 20, comprising: 22. A sample liquid mixture such as blood. Centrifugation in a flexible source container The fractionated components are separated radially relative to the centrifugation axis, and separated during centrifugation. The fractionated components are centrifuged by transferring them from the source container. an apparatus for centrifugal separation, comprising a conduit connected to the source container; During separation, a non-zero centrifugal force gradient is maintained so that the fractionated components in the conduit mix during the transfer. along the length of the mole and radially to the centrifugal axis so as to prohibit A centrifugal separation device arranged to have a directional component directed to. 23. If the conduit is a flexible tube and the device is Claims further comprising a support member for supporting the tube in a radial direction. Apparatus according to clause 22. 24. the conduit is provided with at least one receptacle in front of the conduit in longitudinally spaced portions; at least one claim connected as an inlet to each of the at least one acceptance statement; 24. The device according to paragraph 23. 25. spaced along the conduit to isolate the components to be separated and transported. 25. The apparatus according to claim 24, further comprising means for determining. 26. 26. The device of claim 25, wherein the container is part of a tube. 27. If the isolation means comprises an inner wall of the tube section, said wall is free from centrifugal gradients. sufficient to inhibit mixing of separated components contained along the tube between tube sections. a diameter selected to be small, thereby isolating the separated components; The device according to item 26. 28. The isolation means includes a valve for closing fluid communication to the receiving container. Apparatus according to claim 25. 29. for isolating fluid components contained within sequentially arranged sections of tubing; In this device, there is a stand for supporting the pipe in a straight line near the rail, and a stand on the stand. A rail and a rail to which a plurality of clamp members are slidably attached. , the clamp is configured to attach to the tube to isolate fluid components within the section of the tube. a fluid component capable of being positioned between each of said sequentially arranged portions of the Minute isolation equipment. 30. A clamp forces fluid out of the tube by rolling the clamp over the tube. 30. The apparatus of claim 29, which is a roller clamp for 31. A sample liquid mixture in a source container, such as blood, is centrifuged to separate the fractionated components radially. During centrifugation, the fractionated components are transferred via a conduit to a receiving vessel and The conduit, the source vessel and the receiving vessel form a closed fluid system, and the source vessel is a sample liquid mixture disposed at least partially radially outwardly from said receiving vessel; In a centrifugal separator for separating fractionated components of a compound, the an occluding member for occluding flow through the tube and for allowing or inhibiting flow; means for controlling the closure member to an orifice forming a portion of a flow path from the source vessel to the conduit; In response to the presence of a desired fractionated component in said orifice, said orifice is partially a restraining member for closing the source vessel, thereby reducing the flow rate from the source vessel. Centrifugal separator. 32. The suppression member is a float member, and the float member controls the specific gravity of the fractionated component and its It has a specific gravity between that of the adjacent component and partially closes the orifice. 32. A centrifugal separator according to claim 31, having a shape adapted to be chained. Place. 33. The float member has a specific gravity between that of platelets and neocytes (new red blood cells). 33. The centrifugal separator according to claim 32, comprising: 34. having an outlet for separating fractionated components of a sample fluid mixture such as blood a flexible source container, at least one receiving container, said source container and said at least one receiving container; and one receiving vessel about the axis at a speed sufficient to effect separation of said fractionated components. and means for rotating the centrifugal separator, a conduit extending from the outlet to the at least one receiving vessel to form a closed fluid system; and, a replenishment fluid container; pressurizing the fluid system to prevent the formation of a vacuum in the conduit; A make-up fluid is introduced under pressure into fluid pressure communication with the fluid system to maintain the A centrifugal separator comprising means for 35. the replenishment fluid has a higher density than the density of the fractionated components of the sample mixture; A body container is located radially inward from the source container toward the centrifuge axis and is compensated under pressure. The means for inflowing the replenishment fluid includes a valve provided between the replenishment fluid container and the fluid system. , opening the valve thereby allowing the replenishment fluid to maintain the pressurized state by centrifugal force. Claim 34, further comprising means for causing the fluid to flow into fluid pressure communication. centrifugal separator. 36. The replenishment fluid is a saline solution, and the cell allows the replenishment fluid to flow into the source container. 36. The centrifugal separator according to claim 35. 37. A centrifugal separator is used to separate the desired fractions of a fluid mixture, such as blood. a source vessel and a conductor connected to the source vessel for transferring fluid from the source vessel; a conduit, wherein the desired fractionated component occurs in a characteristic volume within the source vessel; is a tube having a volume at least equal to said characteristic volume, said tube being centrifuged The desired fractionated components are mixed in the tube by coordinating them against the medium centrifugal force gradient. A device that enables transfer without causing any collisions. 38. The diameter of the tube is such that the desired 38. The apparatus of claim 37, which is sufficiently small to prevent mixing of fractionated components. 39. 39. The device of claim 38, wherein the device is less than about 5 mm in diameter. 40. A tube is connected to the source container with an outlet opening, the tube being configured to contain a first desired component. a first portion having a volume at least equal to the characteristic volume adjacent to said outflow opening; and having a characteristic volume of a second desired component located remotely with respect to the outlet opening. a second portion having at least equal volumes; said first and second portions having at least equal volumes; 38. The device of claim 37 having different diameters. 41. Ensure that the desired component has a characteristic volume that is less than about 1% of the volume of the source container. The device according to item 37. 42. the first and second desired components are neocytes and platelets, respectively; 41. The device of claim 40, wherein the second portion has a diameter of less than about 5 mm. 43. A support member for supporting a flexible blood receiving device in a centrifuge. There, Along the centrifugal force gradient of the centrifuge, there is a non-zero component along the length of the conduit. a support member having a spool surface for winding the conduit in a desired configuration; means for determining said winding location to achieve said orientation of the conduit on the surface of the roll; A support member comprising: 44. The means for determining the winding location is the approximate saddle shape of the conduit around the spool surface. The support part according to claim 43, which is provided with a green flange that realizes a shape of winding. Material. 45. Mounted on a flexible source vessel in a defined geometric relationship to the spool surface Claim 44, further comprising a funnel-shaped cap for supporting the source container. supporting member. 46. Allow the conduit to pass through the cap when attaching it to the source container. A slot is provided in the cap to allow the support member to be connected to the integral conduit and 46. A support member according to claim 45, adapted to be mounted on a container.