TWI486442B - Cell culture device and cell culture basket - Google Patents

Description

Cell culture device and carrier cell

The present disclosure relates to a cell culture device, and more particularly to a cell culture device that is easy to observe and culture.

There are many ways to study cells, but the most important thing is to directly observe and analyze individual cells in a group or group. Therefore, cell culture can be said to be the most direct, simplest and most accurate reflection of various activities of life. Can provide a variety of messages that occur in the body. Therefore, cell culture technology has long been an indispensable tool and method for medical, pharmaceutical, and life science research. Especially after the completion of human genome sequencing and stem cell research, cell culture technology has played its value and importance. Sex.

The most mature and widely used cell culture technique at present is the use of a culture dish for planar (2D) culture of cells. The bio-operator loads the cells onto the culture dish and adds the desired culture medium, and then moves each dish to the cell culture incubator for cultivation. When cultured, the cells are attached to the bottom surface of the culture dish, and the culture solution is completely still. Therefore, the nutrients required for cell growth or the metabolic substances produced by the cells are freed by the difference in the concentration of the substances in the liquid. Diffusion exchange.

However, the general 2D culture method will make the cells adhere to the bottom surface of the dish. However, it is impossible to maintain the three-dimensional structure of the cells, and since the culture liquid is stationary and the substance diffuses slowly, these conditions affect the physiological functions of the cells.

Furthermore, the function of the general cell culture incubator is to provide a constant temperature (eg, 37 ° C), constant humidity closed culture environment, so that the culture environment can reach saturated vapor pressure, the humidity can reach 96% or more, to avoid the culture liquid. The evaporation of water evaporates, causing a gradual increase in the concentration of the culture solution to cause cell death, and at the same time, it can stably maintain the carbon dioxide concentration in the air to 5%.

However, the general cell culture incubator is bulky and takes up space, and the equipment is expensive, and it is necessary to place a large number of petri dishes in order to meet the economic benefits, so that even if the number of culture dishes is small, the cell incubator needs to be opened, resulting in energy consumption without Environmental protection.

Furthermore, in a single cell incubator, the environmental conditions of all the culture dishes are all the same, resulting in a non-elastic experimental plan for the cultured cells.

In addition, in recent years, cell culture technology has been based on the concept of bionics, for example, to enable cells to maintain a three-dimensional structure, or even a tissue-like structure, or to use a pump to drive a culture fluid to form a circular perfusion to mimic the human body. Loop and so on.

However, the use of the pump to drive the flow of the culture fluid requires a plurality of interlaced configurations of the pipeline, so that not only the assembly process is large, but also the operation or observation is inconvenient, thereby increasing the probability of failure of the cell culture experiment.

Therefore, the development of a new generation of cell culture equipment that is convenient, easy to use, low-cost, and mass-produced has become a subject that is currently being pursued.

The present disclosure provides a cell culture device which not only does not need to be equipped with a pump to drive the culture solution (having low cost and mass production characteristics), and has a bionic concept (the culture medium is in a flowing state) without a large cell incubator ( With low carbon energy saving effect). In addition, the cell culture device has a simple operation interface, conforms to the operation tools or techniques familiar to general biological experimenters, and is compatible with existing automation devices.

The disclosure is based on the principle of liquid natural convection. Convection usually occurs in flowing liquids and gases (hence the name fluid), while convection, the main way in which fluids propagate thermal energy, is dominated by changes in density caused by temperature differences between different regions of the fluid, resulting in a flow phenomenon. When a gas or liquid is heated, the volume expands and the density decreases, and at the same time, more buoyancy is obtained to rise, and the rising gas or liquid cools and the volume shrinks and the density becomes larger, and the buoyancy decreases and sinks. The sinking gas or liquid will supplement the rising gas or liquid, and the rising gas or liquid will supplement the sinking gas or liquid, thereby forming a circulating state, that is, convection.

Therefore, the cell culture apparatus of the present disclosure utilizes a technique of locally heating the cell culture tank to cause natural convection with a temperature difference in the culture solution, for example, a specific region of the cell culture tank (ie, a cell culture region). Locally warming to a temperature suitable for cell growth (usually about 37 ° C), while in another area (ie, the cooling zone of the culture solution) is reduced to a suitable temperature (such as room temperature), so that the culture solution in the cell culture tank is due to temperature difference Generate a convection loop. Thereby, the flowing culture solution will bring fresh nutrients to the cells quickly and quickly carry away from the cell metabolites, which contributes to the survival rate of the cells, so the invention can be used without any expensive liquid pushing device, such as peristaltic, Various types of pumps, such as syringes, form a biomimetic effect of circulating convection around the cell culture zone.

According to the above description, the present disclosure provides a cell culture device comprising at least one container and a heating unit, the container having a cell placement space and an adjustment space, and the cell placement space and the adjustment space are at least The two channels are in communication, and the heating unit is thermally conducted to the cell placement space, and is not thermally conducted to the adjustment space.

In addition, the cell culture apparatus according to the present disclosure will be described in detail later by way of examples and drawings.

1,2,3,3',4,5,6,7,8,8'‧‧‧ cell culture device

10, 50, 70‧ ‧ containers

10a, 50a‧‧‧

10b, 50b‧‧‧ cover parts

10c‧‧‧Opening Department

100,100’,500‧‧‧ card hole

101‧‧‧Air space

102,702b‧‧‧ openings

11,11’,11”,51,51’‧‧‧ spacer

110,510‧‧‧ block

12,42,52,72‧‧‧heating unit

120‧‧‧ recess

13,13',13",43,53,53',53",73,73',73"‧‧‧ ‧heating unit

20,20’‧‧‧ Carrying cell parts

200‧‧‧ card block

201‧‧‧ Carrying space

30,80‧‧‧pie

31,81‧‧‧Hosting

310‧‧‧Connector

311‧‧‧ gas supply tank

4a‧‧‧Array Module

4b, 8a, 8a’‧‧‧ control module

40‧‧‧Working unit

41‧‧‧ gas supply unit

421‧‧‧First heating department

422‧‧‧second heating unit

431‧‧‧First heat sink

432‧‧‧second heat sink

432”‧‧‧fan

44‧‧‧Insulated channel

500a‧‧‧ bottom

501‧‧‧Insulation Department

51a, 60a‧‧‧ concave structure

51b‧‧‧Funnel structure

60‧‧‧Seat

61‧‧‧ recess

62‧‧‧ recesses

700a, 700b, P1, P2, P1', P2', P3, P4, P5, P6‧‧ channels

701‧‧‧ first trough

701a‧‧‧cell injection

701b‧‧‧ Track

702‧‧‧Second trough

702a‧‧‧ culture solution injection port

703‧‧‧First cover

703a‧‧‧ socket

704‧‧‧Second cover

71,71’‧‧‧ Carrying case

9‧‧‧Microscope

A‧‧‧ cell placement space

B‧‧‧Adjustment space

C‧‧‧ cells

D, d‧‧‧ pipe diameter

F, X, X’, Y, Y’, Z, Z’‧‧‧ flow paths

L‧‧‧ liquid level

R‧‧‧ spacing

S‧‧‧ accommodating space

t,t’,w‧‧‧ gap

1A and 1B are schematic exploded and combined views of the first embodiment of the cell culture apparatus of the present disclosure; wherein the 1A' and 1B' diagrams are different from the 1A and 1B diagrams; FIG. 1D is a schematic side view of another aspect of FIG. 1B; FIG. 2A is a perspective exploded view of another aspect of FIG. 1A; FIG. 2B And FIG. 3A is a schematic exploded view of another aspect of FIG. 2A; FIG. 3A is a perspective exploded view of another aspect of FIG. 2A; FIG. 3B is another aspect of FIG. 3A FIG. 4A is a perspective exploded view of another aspect of FIG. 3A; wherein, FIG. 4A' is a perspective view of another aspect of FIG. 4A, and FIG. 4B is a fourth FIG. Partially lower view, FIG. 4C is a partial top view of FIG. 4A; FIG. 4D is a perspective view of a state of use of FIG. 4A; and FIGS. 5A and 5B are second embodiment of the cell culture apparatus of the present disclosure The three-dimensional decomposition and combination diagram; Figure 5C is a three-dimensional diagram of the operation of Figure 5B (omission of the container); 5C' and 5C" It is a side view of different aspects of Figure 5A; Figures 5D and 5E are partial stereo and side views of different aspects of Figure 5A; Figures 6A and 6B are another state of Figure 5A a three-dimensional decomposition and side view; 6C and 6D are partial stereo and side views of another aspect of the 6A and 6B drawings; and FIGS. 7A and 7B are schematic exploded and combined views of the third embodiment of the cell culture device of the present disclosure; Figure 7C is a side view showing the operation of Figure 7B; Figure 7C' is a partial perspective view of another aspect of Figure 7C; and the 7D and 7D' drawings are partial three-dimensional for the application of Figure 7A. FIG. 8A and FIG. 8B are three-dimensional decomposition and combination diagrams of another aspect of FIG. 7A; FIG. 8A′ is a partial perspective view of another aspect of FIG. 8A; FIGS. 8C and 8C′ FIG. 9 is a perspective view showing the three-dimensional assembly and three-dimensional decomposition of the different application modes of FIG. 8A; FIG. 9 is a perspective view showing the state of use of the third embodiment of the cell culture device according to the disclosure; FIGS. 10A, 10B and 10C are A perspective view of various aspects of the heat dissipating unit of the first embodiment of the cell culture device of the present disclosure; FIGS. 11A, 11B and 11C are various aspects of the heat dissipating unit of the second embodiment of the cell culture device of the present disclosure. Stereoscopic diagram; and figures 12A, 12B and 12C are A perspective view of various aspects of a heat dissipating unit of a third embodiment of the disclosed cell culture device.

The embodiments of the present disclosure are described below by way of specific embodiments, and those skilled in the art can readily appreciate the other advantages and functions of the present disclosure.

It should be noted that the structure, proportion, size, etc. shown in the drawings of this specification are It is intended to be used in conjunction with the disclosure of the specification, and is not intended to limit the scope of the disclosure. It is not technically meaningful, any structural modification or proportional relationship. Changes in size or size, which do not affect the effects and achievable effects of this disclosure, should fall within the scope of the technical disclosure disclosed herein. In the meantime, the terms "upper", "lower", "top", "bottom", "left", "right", "inside", "outside", "first" and "second" are quoted in this manual. And the terms "a" and "the" are used for convenience of description, and are not intended to limit the scope of the disclosure. The change or adjustment of the relative relationship is also considered to be the disclosure. The scope of implementation.

The first embodiment of the cell culture apparatus of the present disclosure is asymmetric. The asymmetric type means that the flow path of the liquid is not symmetrical. (for example, from the A end of the container to the other B end, and then back to the A end, complete a cycle)

As shown in FIGS. 1A and 1B, a cell culture apparatus 1 includes a container 10, a spacer 11 disposed in the container 10, and a heating unit 12.

The container 10 comprises a rectangular trough 10a and a capping member 10b having an unsealing portion 10c. The trough 10a defines a cell placement space (or cell culture zone) A and an adjustment space (or a culture solution cooling). District) B. In the present embodiment, the cover member 10b covers the groove body 10a to form a sealed state of the container 10, so that liquid or gas cannot leak from the cover slit of the cover member 10b, and the unsealing portion 10c is located in the adjustment space. The upper side of B serves as a culture liquid injection port, and the gas can freely enter and exit from the cover slit of the unsealing portion 10c, that is, the gas of a general cell culture dish can freely enter and exit from the cover slit. It should be noted that the shape of the groove body 10a is not limited to a rectangle.

The spacer 11 is a plate shape and is disposed between the cell placement space A and the adjustment space B to isolate the cell placement space A and the adjustment space B and serve as a barrier. For heat transfer, the spacer 11 is joined to the left and right inner side walls of the groove body 10a, and has a slit t between the bottom of the groove body 10a and the cover member 10b, as shown in FIG. 1C, so as to make some gaps. As the upper and lower channels P1 and P2, the cell placement space A and the adjustment space B are connected by two channels P1 and P2.

In the present embodiment, the spacer 11 is solid and integrally formed with the groove body 10a to partition the groove body 10a into the cell placement space A and the adjustment space B; the spacer 11 can also be adhered to the groove body 10a. . Alternatively, as shown in FIG. 1A', the spacer 11' is disposed on the slot 10a in a hanging manner. Specifically, the spacer 11' is provided with a block 110, and the inner side wall of the slot 10a is provided. The card hole 100 suspends the spacer 11' in the slot 10a by the cooperation of the card block 110 and the card hole 100.

Further, in order to avoid direct heat conduction, the solid portion of the spacer 11 or the spacer 11' may be hollowed out to form a hollow spacer 11 or a spacer 11'. Further, air can be used as the spacer 11" for transmitting heat insulation. Specifically, as shown in FIG. 1B', the left and right side walls of the tank 10a are recessed and communicated to form an air space 101 penetrating the left and right side walls. When integrally formed in the production of the tank body 10a, it is integrally formed (left and right molds are integrally molded). In addition, in other aspects, as shown in FIG. 1D, the spacer 11' may also be joined to the bottom of the tank body 10a. And a slit t' is formed between the left and right inner sidewalls of the tank body 10a, so that the slits t' are used as the left and right passages P1', P2'. Specifically, the liquid is left by the spacer 11' The channels P1', P2', and P1 of the square, the right side, and the upper side are circulated, and when the tank body 10a is manufactured, the spacer body 11' may be integrally formed by bulging upward from the bottom of the groove.

The heating unit 12 is thermally conducted to the cell placement space A, and is not thermally conducted to the adjustment space B, so that it can transfer thermal energy to the cell placement space A without being transferred to the adjustment space B. Specifically, the heating unit 12 contacts the cell placement space A, While the adjustment space B is not in contact, heat transfer is performed by means of heat conduction (contact type); in other embodiments, the heat supply unit 12 can also transfer heat energy to the cells by heat radiation or heat convection (non-contact) heating. Place space A.

For use, heat is supplied to the cell placement space A, and the temperature of the liquid in the cell placement space A is increased, for example, the cell placement space A is warmed to about 37 °C. In the present embodiment, the heating unit 12 is a heating plate, which may have a recess 120 to contact the outer surface of the groove body 10a at a position corresponding to the cell placement space A by a locking method.

When the cell culture apparatus 1 is used, as shown in FIG. 1C, the cells (not shown) are placed in the cell placement space A, and then the capping member 10b is placed, and the culture solution is injected from the unsealing portion 10c to the liquid surface L. When the tank body 10a is heated, convection circulation occurs in the liquid in the vessel 10. Specifically, the culture solution located in the cell placement space A is expanded in volume and density is reduced by the heating unit 12, so that more buoyancy is obtained and rises, such as the flow path X, and then enters through the channel P1 above the spacer 11. Adjust space B. When the high temperature culture liquid enters the adjustment space B, it will dissipate heat and cool down, causing the volume to shrink and the density to become larger, and at the same time, the buoyancy is reduced and sinks, such as the flow path Y, and then flows back from the channel P2 below the spacer 11 into the cell placement space A, In order to form a natural convection system. If the culture solution in the cell placement space A has a relatively fast flow rate, the distance between the cell wall of the cell placement space A and the spacer 11 can be reduced, for example, the cell placement space A is smaller than the adjustment space B. The scope.

Furthermore, in the structure shown in Fig. 1D, the high temperature culture solution still enters the adjustment space B from the channel P1 above the spacer 11', and the culture solution after the temperature drop is from the left and right channels P1 of the spacer 11'. ', the bottom of P2' is recirculated into the cell placement space A, such as the flow path F.

Further, preferably, the liquid surface L (as shown in FIG. 1C) exceeds the upper edge of the spacer 11, so that the cell placement space A and the liquid of the adjustment space B can communicate with each other, and the liquid level and the cover member are as close as possible. The smaller the space between 10b (or the top surface of the unsealing portion 10c), the lower the amount of liquid that becomes vapor, while avoiding condensation or dissipation.

In addition, in the cell culture, 5% carbon dioxide gas or oxygen is required, so the gas can be cooled to room temperature (or slightly lower than room temperature), and then the gas is introduced into the tank body 10a from the cover of the unsealing portion 10c ( That is, the space between the liquid surface and the unsealing portion 10c), thereby having the following advantages:

a. The pH of the culture solution and the amount of dissolved oxygen can be adjusted.

b. The cooling conditions of the culture solution can be adjusted to promote natural convection.

c. When steam is generated, the vapor will condense into a liquid state due to the lower temperature air, causing it to drip back into the culture solution in the vessel 10.

As shown in Figs. 2A and 2B, the cell culture device 2 includes a carrier cell member 20 placed in the cell placement space A.

The carrier cell member 20 is made of a scaffold having minute holes, and the stent is in the form of a basket, and the material thereof is, for example, a biocompatible filter membrane or a fenestrated membrane.

In the present embodiment, the carrier cell member 20 is suspended in the cell placement space A of the container 10. Specifically, the carrier cell member 20 is a basket (ie, has a storage space 201 for storing the cells C) and is provided with a card block 200, and a card hole 100' is disposed on the inner side wall of the groove body 10a to borrow the card block 200. The carrier cell member 20 is suspended in the cell placement space A in cooperation with the card hole 100'.

When the cell culture apparatus 2 is used, as shown in Fig. 2B, after the tank body 10a is heated, the liquid in the vessel 10 is convectively circulated. Specifically, the elevated high temperature culture solution A split flow is generated, wherein the flow path Z of one flow passes between the carrier cell member 20 and the groove wall, and the flow path Z' of the other flow flow passes between the carrier cell member 20 and the spacer 11, and then merges, and then passes through the spacer. The channel P1 above the 11 enters the adjustment space B, and is cooled down to the cell placement space A, such as the flow path F.

Furthermore, the design of the carrier cell member 20 in a detachable manner has the following advantages:

1. A fixed spacing r (e.g., 80 um) is maintained between the walls of the carrier cell member 20. When the cell-containing liquid is injected therein, the suspended cells will gradually sink due to gravity to form a cell stack having a width of 80 um. , such as a flaky class organization.

2. Because the thickness of the stacked cell layer and the number of layers are relatively uniform, the nutrients and cell metabolites of the culture solution can be smoothly exchanged for material diffusion. If the cells are free to sink, it is likely to pile up a tissue-like structure with uneven thickness, so that the innermost cells in the thicker cell layer may not be able to obtain sufficient nutrients, or the metabolites cannot be discharged and die.

3. After the cell culture is completed, as long as the carrier cell member 20 is pulled out, other subsequent experiments can be performed. Alternatively, a carrying case (as shown in the 7D or 7D' shown below) in which the culture solution is placed is placed for microscopic observation or short-distance delivery.

Further, in other embodiments, as shown in FIG. 2B', the carrier cell member 20' may be a disk-shaped body and disposed in the cell placement space A, so that when the cell C is injected, the cell C can settle on the carrier cell. The upper surface of the piece 20' is confined between the groove wall of the cell placement space A of the trough 10a and the spacer 11, and the circulating convection medium can enter from below the carrier cell member 20' and flow through the cell C. Surrounded to bring fresh nutrients or take away metabolites.

As shown in FIG. 3A, the container 10 can also communicate with the outside world in a gas-like manner, that is, the adjustment space B is connected to the outside by the gas type, and thus the capping member 10b can be formed without forming the unsealing portion 10c. Specifically, the container 10 (or the tank body 10a) is formed with an opening 102 at the bottom of the adjustment space B as a gas exchange port, and the sealing body 102 is covered with a piece of body 30 so that liquid does not leak from the joint.

In the present embodiment, the sheet body 30 is made of a gas permeable material, which allows oxygen or carbon dioxide gas to pass freely, but blocks liquid leakage. Thereby, the cell culture device 3 has the following advantages:

1. After the culture solution is heated, water vapor is generated, and the gas must be collected at the upper side. Therefore, the sealing cover member 10b prevents the water vapor from overflowing, so that the concentration of the culture solution does not become high.

2. Since the container 10 is sealed above and below, the contamination of bacteria or mold can be completely isolated.

3. When the cells are cultured, the required 5% carbon dioxide gas or oxygen will enter through the sheet 30 to adjust the pH and dissolved oxygen of the culture solution, and also effectively filter impurities such as bacteria, mold or mold in the input gas. spore.

4. A large area of opening 102 can be designed to increase the exchange of gases in accordance with the range of the adjustment space B.

5. The tank body 10a can also design a gas exchange port on the side wall of the adjustment space B to increase gas exchange.

Further, in other aspects, the capping member 10b may also form the unsealing portion 10c, but the unsealing portion 10c is sealed so that liquid or gas cannot freely enter and exit from the slit of the unsealing portion 10c.

Further, the cell culture device 3 includes a carrier 31 for carrying the container 10, As the air supply unit, the carrier 31 has a gas supply groove 311 corresponding to the opening 102 (to connect the gas source to the port 310), and the gas enters the adjustment space B via the sheet 30. When the container 10 is placed on the carrier 31, a sealed space is formed which does not leak gas.

As shown in FIG. 3B, the cell culture device 3' has a plurality of containers 10 (containing a spacer). When a large number of cells are cultured, a plurality of carriers 31 can be connected in series, so that the containers 10 are arranged in series, and Each of the carriers 31 is electrically connected to each other in a communicating pipe manner (that is, the respective ports 310 are communicated with each other) to provide a gas having stable and uniform composition conditions (e.g., concentration).

In this embodiment, each of the carriers 31 can also be designed to be independent, that is, each of the ports 310 can transport the same or different gases.

Furthermore, the heating units 12 can be connected to provide a uniform temperature; or, the heating units 12 can be separated to set different temperatures or share a heating controller (ie, the temperatures of the heating units 12 are the same).

Further, the respective containers 10 are indirectly electrically connected to each other by the carrier 31.

As shown in Fig. 4A, when a large number of cells are cultured, the containers can be arranged in an array. Specifically, an array type module 4a composed of a plurality of working units 40 (24 in the figure) and a control module 4b corresponding to the array type module 4a are provided.

The working unit 40 includes a tank body 10a, a partition body 11, a body 30, and the like, and the control module 4b includes a gas supply unit 41, a heat supply unit 42 and a heat dissipation unit 43, and the control module 4b The outer shape completely matches the bottom structure of the array type module 4a, so that the array type module 4a can be directly heated, cooled, and supplied with air.

Furthermore, the sheet body 30 can be attached to the bottom of the tank body 10a when the array type module 4a is manufactured. The cover member 10b of the array type module 4a can be designed as a separate 24 or a single large panel cover. The carrier-carrying member 20 of the array type module 4a can be used to manufacture an array The module 4a is placed or placed in use.

The heating unit 42 is thermally conducted to the cell placement space A without being thermally conducted to the adjustment space B. Specifically, the heating unit 42 is divided into a first heating unit 421 and a second heating unit 422, and the first heating unit 421 is disposed on the upper portion of the control module 4b, and the second heating unit 422 is disposed in the control module. Below the 4b.

The heat radiating unit 43 is thermally conducted to the adjustment space B, and is not thermally conducted to the cell placement space A. Specifically, the heat dissipation unit 43 is divided into a first heat dissipation portion 431 and a second heat dissipation portion 432. The first heat dissipation portion 431 is a temperature reduction plate and is disposed on the upper portion of the control module 4b, and the second heat dissipation portion 432 is, for example, heat dissipation. The fins (or a fan 432 may be attached to one side of the heat sink fins) to accelerate the temperature reduction, thereby promoting natural convection of the culture solution, as shown in FIG. 4A', and disposed at the lower portion of the control module 4b.

In the present embodiment, the first heating portion 421 and the first heat dissipating portion 431 form an accommodation space S to correspondingly position the trough 10a. Preferably, as shown in FIG. 4B, the groove body 10a protrudes from the bottom side of the array type module 4a to facilitate the insertion into the accommodating space S.

Furthermore, the second heating portion 422 of the heating unit 42 and the second heat dissipating portion 432 of the heat dissipating unit 43 may form a heat insulating structure, such as a heat-insulating material, an air-type heat insulating passage 44, or the heat insulating passage 44. Fill the heat-resistant material in the middle. The heat energy of the second heating portion 422 of the heating unit 42 is prevented from being quickly transmitted to the second heat dissipating portion 432 of the heat dissipating unit 43 by the air or the heat resisting material, so that the temperature difference is reduced, thereby affecting the convection effect, and the two can be avoided. Produces heat transfer.

Moreover, the control module 4b includes two sets of heating units 42 and three sets of heat dissipating units 43 which are staggered with each other, and each heating unit 42 heats two rows of working units 40 at the same time. Specifically, if any two rows of the troughs 10a are arranged in a cell placement space A as a face to face, a group of heating units 42 can be shared, thereby reducing the array. The volume of the column module 4a and the control module 4b. As shown in Fig. 4A, the cell placement space A of the trough 10a of the first row is adjacent to the cell placement space A of the trough 10a of the second row, and the cell placement space of the trough 10a of the third row is as shown. A is adjacent to the cell placement space A of the tank 10a of the fourth row. Moreover, the heat dissipating units 43 on both sides radiate only one row of the working unit 40, and the heat dissipating unit 43 in the middle simultaneously dissipates the two rows of the working units 40.

The air supply unit 41 communicates with each of the tank bodies 10a in a gas-like manner. Specifically, the air supply unit 41 is designed to form a communication tube structure by using the serial carrier 31 as a row, and is disposed under the control module 4b, as shown in FIG. 4C, and the bottom of the accommodation space S The gas supply tank 311 is provided to allow gas to enter the respective tank bodies 10a via the sheets 30.

In the present embodiment, the control module 4b has four sets of air supply units 41, so that the four rows of work units 40 can be supplied with different gas components, or all the same components. It is also possible to design only one set of gas supply units 41 to supply all of the work units 40 in a single line.

When the cell culture device 4 of the array modular structure is used to observe the cell state, as shown in FIG. 4D, the array module 4a can be easily taken out from the control module 4b, and the array module 4a can be directly placed. On the microscope 9, the entire batch of cells was observed from the bottom of the vessel 10. After viewing, the cells are again returned to the control module 4b for further cell culture.

The second embodiment of the cell culture device of the present disclosure is a symmetrical type, wherein the symmetric flow means that the flow paths of the liquid are symmetrical (for example, from the center of the bottom of the container to the top end, the radiation flows out in all directions, Then return to the center of the bottom along the inner wall of the container to complete a cycle). The main difference between this embodiment and the above embodiment lies in the shape of the spacer and the position of heating.

The cell culture device 5 shown in Figures 5A and 5B has a container 50 containing one The circular barrel-shaped tank body 50a and a cover member 50b covering the tank body 50a, and the cell placement space A is an intermediate circle and a projected area thereof (defined by the large diameter D area of the spacer 51) ), and the adjustment space B is the outer ring and the area of its projection (ie, the defined range of the imaginary line). It should be noted that the shape of the groove body 50a is not limited to the shape of a barrel.

Further, the cover member 50b is covered with the groove body 50a so that liquid or gas can leak from the cover slit of the cover member 50b. Alternatively, the structure of the unsealing portion 10c as shown in Fig. 1A can be designed.

The heating unit 52 is supported in a columnar manner to support the bottom of the tank body 50a, and contacts the outer surface of the bottom of the tank body 50a at a position corresponding to the cell placement space A to heat the cell placement space A to about 37. °C.

The spacer 51 is a solid tubular shape and has a concave-convex structure 51a at its bottom portion to contact the inner bottom surface of the groove body 50a, so that the uneven structure 51a serves as a plurality of channels P3 that connect the cell placement space A and the adjustment space B. Alternatively, the spacer 51 may be suspended, that is, it does not contact the inner bottom surface of the groove body 50a, so that the uneven structure 51a is not required to be formed.

When the cell culture apparatus 5 is used, as shown in Fig. 5C, cells (not shown) are first placed in the cell placement space A, and when the tank body 50a is heated, convection circulation occurs in the liquid in the container 50. Specifically, the culture liquid located in the cell placement space A is expanded in volume and density is reduced by the heating unit 52, so that more buoyancy is obtained and rises, and then enters the adjustment space B via the channel P4 in the middle of the spacer 51, such as Flow path X'. When the high temperature culture liquid enters the adjustment space B, it will dissipate heat and cool down, causing the volume to shrink and the density to become larger, and at the same time, the buoyancy is reduced and sinking, and then flows back from the channel P3 below the spacer 51 into the cell placement space A, such as the flow path Y' In order to form a natural convection system. Therefore, the liquid system flows inward or outward in a radial direction, that is, symmetric.

In the present embodiment, the spacer 51 is provided on the groove body 50a in a hanging manner. Specifically, the spacer 51 is provided with a card block 510, and the inner side wall of the slot body 50a is provided with a card hole 500 for engaging the spacer 51 in the container 50 by the cooperation of the card block 510 and the card hole 500.

Further, it is also possible to use air as a spacer, that is, the spacer 51' is hollow, and as shown in Fig. 5C', it is more conducive to the transmission of heat.

Further, in order to allow the culture medium in which the cells are placed in the space A to have a relatively fast flow rate, the range of the cell placement space A may be inconsistent, and as shown in Fig. 5C", the diameter d of the spacer 51 is reduced. The spacer 51 may have a funnel structure 51b, such as a bottom portion of the cell placement space A, which is tapered so that the culture liquid can smoothly flow into the small pipe. Therefore, when the range of the cell placement space A is reduced, the borrowing can be borrowed. The liquid is directed by the funnel structure 51b toward the intermediate conduit.

Further, as shown in FIGS. 5D and 5E, the bottom portion 500a of the tank body 50a has a heat insulating portion 501 which is located between the cell placement space A and the adjustment space B to prevent heat from passing through the bottom of the tank body 50a. 500a is conducted to the adjustment space B, thereby isolating the heat conduction of the material. Specifically, the bottom portion 500a of the groove body 50a is removed from the material between the cell placement space A and the adjustment space B, so as to form a channel on the outer surface of the bottom portion 500a of the groove body 50a as the heat insulating portion 501, thereby thinning The thickness of the bottom of the tank body 50a (ie, the wall thickness between the connected cell placement space A and the adjustment space B is thin), so as to prevent the heat of the intermediate cell placement space A from being transmitted to the peripheral adjustment space B through the groove wall, And affect the effect of convection.

As shown in Figs. 6A to 6D, in another aspect of the cell culture device 6, the container 50 is provided with a boss 60, and the spacer 51 has a recess 61 corresponding to the boss 60 to form a funnel structure.

As shown in Figures 6A and 6B, the platform portion of the boss 60 is used to place cells, A pocket 62 is formed on the outer surface of the bottom of the tank body 50a for the heat supply unit 52 to be inserted into the pocket 62. Specifically, when the tank body 50a is manufactured, the boss 60 and the pocket 62 are integrally formed.

Further, when the spacer 51 is placed in the groove body 50a, the spacer 51 is suspended, and a slit w is formed between the boss 60 and the recess 61 as a passage P5 of the liquid flow path F. Preferably, the spacer 51 adopts a small diameter d and cooperates with a tapered funnel structure 51b.

Further, as shown in FIGS. 6C and 6D, the end surface of the projection 60 may have a concave-convex structure 60a such as a groove, and the concave-convex structure 60a may contact the surface of the concave portion 61 of the spacer 51 to form the concave-convex structure 60a and the concave portion 61. The slit serves as a passage P6 of the flow path F of the liquid.

In addition, in the second embodiment, the design of the cover member, the sheet body, the carrier, the serial connection, the array module, and the like can be designed with reference to the technical means of the first embodiment.

The third embodiment of the cell culture device of the present disclosure is a loop-shaped cassette type, the ring shape refers to the appearance of the container, and the cassette type refers to the overall appearance. The main difference between this embodiment and the above embodiment is that no spacers need to be provided.

As shown in FIGS. 7A, 7B, and 7C, the cell culture device 7 includes a container 70 and a heat supply unit 72.

The container 70 includes a first tank body 701, a second tank body 702, a first cover member 703, a second cover member 704 and two passages 700a, 700b. The first tank body 701 is defined as a cell placement space A. The second tank body 702 is defined as an adjustment space B, and the two channels 700a, 700b are respectively disposed above and below the container 70, and the first and second cover members 703, 704 are respectively provided with the first and second tank bodies. 701,702. Since the cell placement space A is completely separated from the adjustment space B, there is a good thermal insulation effect between the two.

The first tank body 701 has a cell injection port 701a above it for direct injection into the fine The cell, or can be placed directly into the carrier cell 20 . Preferably, the width of the first tank 701 can be reduced (for example, the volume of the first tank 701 is smaller than the volume of the second tank 702) to accelerate the flow rate of the culture solution.

Above the second tank 702, there is a culture liquid injection port 702a for directly injecting the culture solution or replacing the medicine.

The first cover member 703 is provided with a cell injection port 701a and is tightly sealed so as not to leak liquid or gas.

The second cover member 704 covers the culture liquid injection port 702a and is not in close contact so that the gas can freely enter and exit from the cover slit.

The heating unit 72 is configured to chuck the outer surface of the first tank body 701 to heat the liquid in the first tank body 701 to about 37 ° C, and generate convection by the two passages 700a, 700b, as shown in FIG. 7C. Path X, Y.

In the present embodiment, the carrier cell member 20 is placed in the first tank body 701. Specifically, as shown in FIG. 7C (see also FIG. 2A), a rail 701b is provided in the first tank body 701 to engage the carrier cell member 20 with the first tank body 701.

Alternatively, as shown in FIG. 7C', the carrier cell member 20 is inserted into the first cover member 703 (ie, inserted into the socket 703a), so that when the first cover member 703 is disposed on the first slot body 701, The carrier cell member 20 is positioned in the cell placement space A. In this way, the carrier member 20 and the first cover member 703 can be combined before delivery, so that the user can only easily insert and insert the cell cover space A and the seventh cover member 703 by contacting the first cover member 703. The carrying case 71 is shown in the figure without contaminating the carrier cell member 20.

Furthermore, the carrier cell member 20 can be of various portable designs. The carrying case 71 shown in FIG. 7D is designed according to the first slot body 701 shown in FIG. 7A, or as The carrying case 71' shown in Fig. 7D' is designed in accordance with the first groove 701 having the rail 701b shown in Fig. 7C.

In addition, in the cell culture, 5% carbon dioxide gas or oxygen is required, so the gas can be first cooled to room temperature (or slightly lower than room temperature), and then the gas is injected from the second capping member 704 and the culture solution into the port 702a. The gap between the gaps enters the second tank 702.

In another aspect, the cell culture device 8 shown in Figs. 8A and 8B has a container 70 that communicates with the outside in a gas-like manner. Specifically, the bottom of the second tank 702 has an opening 702b, and the opening 702b is covered by a piece 80.

Further, the cell culture device 8 includes a carrier 81 for supplying gas. For technical details of the sheet 80 and the carrier 81, reference is made to the description of FIG. 3A.

Moreover, the carrier 81 can be used as a gas supply unit to integrate the heating unit 72 (and the heat dissipation unit 73), such as heat dissipation fins, into the control modules 8a, 8a' as shown in Figs. 8A and 8A'. Preferably, when the container 70 is placed on the control module 8a', the outer wall surface of the second tank 702 contacts the heat dissipating unit 73".

In another aspect, as shown in FIG. 8C, a plurality of control modules 8a may be connected in series, whereby when the cell culture device 8' has a plurality of containers 70, the containers 70 are arranged in series. Each of the control modules 8a is electrically connected to each other in a communication tube manner, that is, a gas having the same composition is supplied to each of the containers 70.

Alternatively, as shown in Fig. 8C', the control modules 8a are arranged side by side, that is, each control module 8a is separately supplied with gas to each of the containers 70 to supply different (or the same) gases to the respective containers 70.

In addition, each control module 8a can be designed to operate independently, that is, each heating unit 72 is independently controlled, so that different temperatures can be set, or a heating controller can be shared, so that the temperatures of the respective heating units 72 are the same.

As shown in Fig. 9, in the cell culture apparatus 7, 8, 8' of the third embodiment, the container 70 was placed directly on the microscope 9 to observe the cells. After viewing, the cell culture can be continued by returning to the control unit 8a.

Various aspects of the heat dissipating unit that can be designed in the first to third embodiments will be described below.

As shown in FIGS. 10A, 11A and 12A, the heat dissipating units 13, 53, 73 are designed as jagged, wavy sidewalls of the adjustment space wall structure of the container to increase the heat dissipation area.

As shown in Figures 10B, 11B and 12B, the heat dissipating units 13', 53', 73' are planar heat dissipating blocks of a highly thermally conductive material such as metal or glass to directly contact the outer surface of the adjustment space of the container.

As shown in Figures 10C, 11C and 12C, the heat dissipating units 13", 53", 73" are heat-dissipating fins of high thermal conductivity materials such as metal and glass to directly contact the outer surface of the adjustment space of the container. .

In addition, the heat sink wall design can be combined with the heat sink block (or heat sink fin).

In summary, the cell culture device of the present invention mainly comprises a biomimetic effect by dividing the container into a cell placement space and an adjustment space, and heating the cell placement space to generate natural convection and accelerate material exchange. And can achieve good cell culture efficiency.

Furthermore, only the cells are placed in a space for heating, which not only consumes low power, but also is environmentally friendly and energy-saving.

Moreover, when a large number of cells are cultured, the experiment can be achieved by modularizing to meet the demand for miniaturization, and different parameter conditions can be set according to requirements, and the cells are cultured.

In addition, by carrying the cell parts, the cells can be stacked into a cell stack with uniform thickness (similar to tissue slices, with biomimetic effect) to maintain cell-to-cell contact and maintain the three-dimensional structure of the cells, thus helping The physiological performance of the cells and the long-term culture.

The above embodiments are intended to illustrate the principles of the disclosure and its functions, and are not intended to limit the disclosure. Any person skilled in the art can modify the above embodiments without departing from the spirit and scope of the disclosure. Therefore, the scope of protection of the present disclosure should be as set forth in the scope of the patent application described later.

1‧‧‧ cell culture device

10‧‧‧ Container

10a‧‧‧Slot

10b‧‧‧Cover parts

10c‧‧‧Opening Department

11‧‧‧ spacer

12‧‧‧heating unit

A‧‧‧ cell placement space

B‧‧‧Adjustment space

P1, P2‧‧ channel

X, Y‧‧‧ flow path

L‧‧‧ liquid level

T‧‧‧ gap

Claims (18)

A cell culture device comprising: at least one container having a cell placement space and an adjustment space, wherein the cell placement space and the adjustment space are connected by at least two channels; and the heating unit is paired The cells should be placed in a space, and the heating unit directly thermally conducts the cell placement space. The cell culture device according to claim 1, wherein the container is provided with a spacer to distinguish the cell placement space from the adjustment space, and the spacer has a gap with the container. Take this channel. The cell culture device according to claim 2, wherein the spacer system is solid or hollow. The cell culture device of claim 2, wherein the spacer system has an air space. The cell culture device according to claim 2, wherein the spacer system is plate-shaped or tubular. The cell culture apparatus according to claim 2, wherein the spacer system has a concave-convex structure, and the concave-convex structure contacts an inner surface of the container. The cell culture apparatus according to claim 2, wherein the container is provided with a boss, and the spacer system has a recess corresponding to the boss. The cell culture apparatus according to claim 2, wherein the container has a concavo-convex structure, and the concavo-convex structure contacts the spacer. The cell culture apparatus according to claim 1, wherein the container is connected to the outside by a gas type. The cell culture device according to claim 9 of the patent application, further comprising a gas supply list A unit that communicates the container in a gas-like manner. The cell culture apparatus according to claim 1, wherein when the container has a plurality of containers, each of the containers is electrically connected to each other. The cell culture apparatus according to claim 1, wherein when the container has a plurality of containers, each of the containers is arranged in series or in an array. The cell culture apparatus according to claim 1, wherein the container has a heat insulating portion between the cell placement space and the adjustment space. The cell culture device according to claim 1, wherein the volume of the cell placement space is smaller than the volume of the adjustment space. The cell culture device according to claim 1, further comprising a carrier cell member placed in the cell placement space. The cell culture device according to claim 15, wherein the material for carrying the cell member is made of a stent having minute holes. The cell culture device according to claim 16, wherein the stent has a biocompatible filtration membrane or a porous membrane. The cell culture device according to claim 1, further comprising a heat dissipating unit disposed to adjust the space, and the heat dissipating unit directly thermally conducts the adjusting space.