CN110106080B

CN110106080B - Cell separation device

Info

Publication number: CN110106080B
Application number: CN201910502927.8A
Authority: CN
Inventors: 牛海涛; 凌健东; 管志平; 许爱红; 杨雪; 秦晓瑞
Original assignee: Shenzhen Haisian Biotechnology Co ltd
Current assignee: Shenzhen Haisian Biotechnology Co ltd
Priority date: 2019-06-11
Filing date: 2019-06-11
Publication date: 2024-08-02
Anticipated expiration: 2039-06-11
Also published as: CN110106080A

Abstract

The application discloses a cell separation device which comprises a separation seat, at least one separation cup and at least one magnet, wherein the separation cup is used for containing cell mixed liquid with magnetic beads, the separation cup is detachably arranged on the separation seat, the magnet is arranged on the separation seat, and the magnet is arranged below the separation cup and is used for adsorbing the magnetic beads to the bottom of the separation cup. The cell separation device has the advantages of simple structure, low cost and high separation efficiency.

Description

Cell separation device

Technical Field

The application relates to the technical field of biology, in particular to a cell separation device.

Background

Lymphocytes (lymphocytes) are one of the white blood cells, the smallest volume of which is produced by lymphoid organs and which are important cellular components of the immune response function of the body. Lymphocytes are a type of cell line with immune recognition function, and can be classified into T lymphocytes (also called T cells), B lymphocytes (also called B cells), natural Killer (NK) cells, etc., according to their migration, surface molecules, and functions. T cells and B cells are antigen-specific lymphocytes, which are initially identical in origin, and are derived from hematopoietic tissues. T lymphocytes circulate with blood to the thymus, mature under the action of thymus hormones and the like, while B cells differentiate and mature in the bone marrow.

When stimulated by antigen, T lymphocyte is converted into lymphoblast, and then is subdivided into sensitized T lymphocyte, and the sensitized T lymphocyte participates in cell immunity, and the immune function is mainly resisting intracellular infection, tumor cell, allogeneic cell and the like; b lymphocytes are transformed into plasmablasts, and then differentiated into plasma cells, and produce and secrete immunoglobulins (antibodies) to participate in humoral immunity, which function to produce antibodies, present antigens, and secrete intracellular factors to participate in immunomodulation; NK cells exert spontaneously cytotoxic effects independent of antigen stimulation, and have the effect of killing target cells.

Twenty years ago, with the advent of recombinant hormone, soluble receptor and antibody based biological agents, an important revolution has emerged in the pharmaceutical industry under the control of small molecule drugs as support for a long time. Now, along with the development of the multipotency of the treatment of microorganisms and human cells, biological medicine is pushed to the blast tip, and a new revolution is initiated. Continuous advances in cell engineering provide a systematic framework for developing safe, predictable cell therapies. Microorganisms and human cells are also used as therapeutic entities, potentially addressing some important, currently unmet needs, for the treatment of certain most deadly diseases, including cancer, autoimmune diseases, and the like.

Researchers at the university of california, san francisco, in the journal of science-transformation medicine (ScienceTranslationalMedicine), column PERSPECTIVE, published an article on the overview of the prospect of cell therapy. The article states that cell therapy will become the "third largest medical stay" in the future, and will be used universally to treat patients as drugs now made with engineered proteins, antibodies or smaller chemicals. Researchers have come to the metaphor that if small molecules and biological products are tools, then the cells are carpenters, architects, and engineers. The cells can perform some functions that cannot be achieved by small molecule drugs and targeted drugs. For example, cells are adaptable, they can migrate to specific sites, sense the surrounding environment better than current drugs, then make their correct decisions themselves, change their responses, and adapt better to physiological conditions.

First, cells are naturally responsible for many therapeutic tasks, such as phagocytosis of pathogens by macrophages, recruitment of adaptive immune cells, production of myeloid and lymphoid lineage cells by hematopoietic stem cells, production of chondrocyte extracellular matrix by chondrocytes, and the like.

Second, the behavior of the cells is selective. For small molecules and biological agents, they do not have a switch, and exert their biological effects as long as they bind to the target. However, cells are able to sense the surrounding environment and only act when a specific signaling molecule is triggered. Therefore, the cell therapy can better avoid off-target effect, and also can play a better delivery role, thus being a good carrier for other therapies.

Third, cell therapy can better adapt to human genetic diversity. For example, due to individual differences, drugs are metabolized differently in the human body and the therapeutic effects are also different. But through reforming cells, the concentration change can be automatically regulated like a resistance circuit, and the cell is suitable for metabolism of different hosts, so that better curative effect is exerted.

Finally, the function of the cells can be regulated by altering the cellular genes. For example, T lymphocytes are modified by a biosynthesis technology, so that the T lymphocytes can sense blood sugar and secrete insulin, and the dependence of type 1 diabetics on blood sugar is relieved; immune responses against cancer are often weaker, but by manipulating and incubating populations of immune cells that target specific molecules on cancer cells, anti-tumor responses can be enhanced. The research of the new experiments is a great breakthrough for clinical treatment.

With the continued maturation of cytotherapeutic techniques, mass lymphocyte separation is one of the key elements.

At present, the cell separation technology is mature and has wider application mainly as follows: density gradient centrifugation, immunomagnetic bead methods, flow techniques, and the like. The density gradient centrifugation method is simple to operate and low in cost, but can only roughly separate cell populations, and cannot meet the requirement of complex cell separation in clinical and scientific research; the immunomagnetic bead method and the flow cell separation method are both based on the principle of capturing cells by antibodies, utilize specific markers on the cell surface and combine with other technologies to realize specific cell separation, and the cells separated by the two technologies can be specific subgroups, so that the purity is relatively high.

The magnetic beads used for separating cells by immunomagnetic bead method comprise large magnetic beads (0.1-0.45 mm) and small magnetic beads (about 50-5000 nm). The existing method for separating cells by using large magnetic beads on the market has the defects of simple operation, high separation speed, low cost and low purity, and causes mechanical pressure on the cells, thereby influencing the biological activity of the cells, being unfavorable for culturing after separation, and the separated large magnetic beads must separate the cells from the magnetic beads by a certain method; the method for separating cells by using the small magnetic beads basically adopts a separation column, has high separation purity and good cell activity, and the magnetic beads can directly flow upwards without shearing; but the separation speed is low, the separation column is disposable, the cost is high, the separation column is required to be eluted with negative cells and positive cells after passing through the column, and the operation is complicated.

The existing cell separation device has the problems of complex structure, high cost and low separation speed.

Disclosure of Invention

The application provides a cell separation device, which aims to solve the problems of complex structure, high cost and low separation speed of the cell separation device in the prior art.

In order to solve the above problems, the present application provides a cell separation device, which comprises a separation seat, at least one separation cup and at least one magnet, wherein the separation cup is used for accommodating a cell mixed solution with magnetic beads, the separation cup is detachably arranged on the separation seat, the magnet is arranged on the separation seat, and the magnet is arranged below the separation cup and is used for adsorbing the magnetic beads to the bottom of the separation cup.

Wherein, the longitudinal section of the separating cup is V-shaped.

Wherein, two adjacent separation cups combine to form a separation cup assembly, and the separation cup assembly is integrally formed.

The separating seat comprises a partition plate, a plurality of accommodating grooves are formed below the partition plate and are used for accommodating magnets, a guide plate assembly perpendicular to the partition plate is arranged above the partition plate, and the guide plate assembly comprises a first guide plate and a second guide plate which are arranged in parallel at intervals; the separating cup assembly comprises a first separating cup, a second separating cup and a connecting block for connecting the first separating cup and the second separating cup, the connecting block is of an inverted V shape, a first clamping groove and a second clamping groove are formed in the connecting position of the connecting block and the first separating cup and the connecting position of the connecting block and the second separating cup respectively, the first clamping groove is clamped with the first guide plate, and the second clamping groove is clamped with the second guide plate.

The separating seat further comprises a side plate, the side plate is arranged on one side of the partition plate and perpendicular to the partition plate and the guide plate assembly, a limiting hole is formed in the side plate, the connecting block is outwards protruded to be provided with a limiting rod, and the limiting hole is matched with the limiting rod.

The side surfaces of the separating cup assemblies are all in a plane shape, and the bottoms of the first separating cup and the second separating cup are in a plane shape to be abutted with the partition plate.

The upper end faces of the first guide plate and the second guide plate are subjected to round corner guiding treatment, and the first clamping groove and the second clamping groove are subjected to round corner guiding treatment.

The separating seat further comprises a packaging plate, wherein the packaging plate is parallel to the side plate and used for packaging the magnet.

Wherein, adjacent three separation cups combine to form separation cup subassembly, separation cup subassembly integrated into one piece.

The separating seat comprises a partition plate, a plurality of accommodating grooves are formed below the partition plate and are used for accommodating magnets, a first guide plate assembly and a second guide plate assembly which are perpendicular to the partition plate are arranged above the partition plate, the first guide plate assembly comprises a first guide plate and a second guide plate which are arranged at intervals in parallel, and the second guide plate assembly comprises a third guide plate and a fourth guide plate which are arranged at intervals in parallel; the separating cup assembly comprises a first separating cup, a second separating cup, a third separating cup, a first connecting block and a second connecting block, wherein the first connecting block is connected between the first separating cup and the second separating cup, the second connecting block is connected between the second separating cup and the third separating cup, the first connecting block and the second connecting block are of inverted V shapes, a first clamping groove and a second clamping groove are formed in the connection position of the first connecting block and the first separating cup and the second separating cup respectively, the first clamping groove is in clamping connection with the first guide plate, the second clamping groove is in clamping connection with the second guide plate, a third clamping groove and a fourth clamping groove are formed in the connection position of the second connecting block and the second separating cup and are respectively in clamping connection with the third guide plate, and the third clamping groove is in clamping connection with the third guide plate and the fourth clamping groove is in clamping connection with the fourth guide plate.

The cell separation device comprises a separation seat, a separation cup and a magnet, wherein the separation seat can be reused, and target cells in the separation cup can be adsorbed by the magnet, so that cell separation is realized. The cell separation device has the advantages of simple structure, low cost and high separation efficiency.

Drawings

FIG. 1 is a schematic view showing the structure of an embodiment of a cell separation apparatus according to the present application;

FIG. 2 is a schematic diagram showing the structure of the cell separation apparatus shown in FIG. 1 at another view angle;

FIG. 3 is a schematic view of the structure of an embodiment of a separation cup assembly of the cell separation apparatus shown in FIG. 1;

FIG. 4 is a schematic flow chart of a first embodiment of the cell separation method of the present application.

Detailed Description

The cell separation device comprises a separation seat, at least one separation cup and at least one magnet, wherein the separation cup is used for containing cell mixed liquid with magnetic beads, the separation cup is detachably arranged on the separation seat, and the magnet is arranged below the separation cup and is used for adsorbing the magnetic beads to the bottom of the separation cup. The cell mixture solution with magnetic beads may be a magnetic bead and a cell sample solution to which an antibody corresponding to a target cell is coupled, or in other embodiments, a magnetic bead and a cell sample solution to which a substance capable of specifically binding to a target cell is coupled. The separating cup can enable the magnetic beads adsorbed with the target cells to be firmly adsorbed at the bottom of the separating cup without wall hanging, so that the yield is improved.

The cell separation device comprises a separation seat, a separation cup and a magnet, wherein the separation seat can be reused, the separation cup is preferably disposable, and target cells in the separation cup can be adsorbed by the magnet, so that cell separation is realized. The cell separation device has the advantages of simple structure, low cost and high separation efficiency.

Referring to fig. 1-3, fig. 1 is a schematic diagram illustrating a cell separation apparatus according to an embodiment of the application; FIG. 2 is a schematic diagram showing the structure of the cell separation apparatus shown in FIG. 1 at another view angle; FIG. 3 is a schematic view of the structure of an embodiment of a separation cup assembly of the cell separation apparatus shown in FIG. 1. In this embodiment, the cell separation apparatus includes a separation housing 10, a separation cup assembly 20, and a magnet 30.

The separating seat 10 comprises a partition plate 101, a side plate 102 and a packaging plate 103, the partition plate 101 is a horizontal plate body, a plurality of accommodating grooves 104 are formed below the partition plate 101 and are used for accommodating the magnets 30, the plurality of accommodating grooves 104 are arranged at intervals along the length direction of the partition plate 101, and each magnet 30 corresponds to a separating cup of the separating cup assembly 20. Preferably, the accommodating groove 104 is square, the magnet 30 is a square magnet 30, and is a neodymium iron boron with galvanized surface, grade N35, and size 7mm×10mm. Preferably, the separating seat 10 is formed of a white ABS material.

A guide plate assembly perpendicular to the partition plate 101 is arranged above the partition plate 101, and comprises a first guide plate 105 and a second guide plate 106 which are arranged in parallel at intervals, and the guide plate assembly comprises a first guide plate 105 and a second guide plate 106 which are arranged in parallel at intervals.

The side plate 102 is arranged on one side of the partition plate 101 and is perpendicular to the partition plate 101 and the guide plate assembly, and a limiting hole 107 is formed in the side plate 102.

The package plate 103 is parallel to the side plate 102 for packaging the magnet 30. After the magnet 30 is placed in the accommodating groove 104, the packaging plate 103 can be used for sealing, so that the magnet 30 is prevented from falling out, and the packaging plate 103 can be adhered or clamped on one surface of the opening of the accommodating groove 104.

The separation cup assembly 20 is used for containing cell mixed liquid with magnetic beads, the separation cup assembly 20 is detachably arranged on the separation seat 10, and the magnets 30 are in one-to-one correspondence with the separation cups at the bottom of the separation cup assembly 20. The cell mixture solution with magnetic beads may be magnetic beads and cell sample solution, to which antibodies corresponding to target cells are coupled, and the diameters of the magnetic beads are preferably 500-5000 nm, and in other embodiments, magnetic beads and cell sample solution to which substances capable of specifically binding to target cells are coupled may be used. The separation cup assembly 20 can enable the magnetic beads adsorbed with the target cells to be firmly adsorbed at the bottom of the separation cup assembly 20 without hanging on the wall, thereby improving the yield. Specifically, the split cup assembly 20 is formed by combining two adjacent split cups, and the split cup assembly 20 is preferably integrally formed. The separating cup assembly 20 comprises a first separating cup 201, a second separating cup 202 and a connecting block 203 for connecting the first separating cup 201 and the second separating cup 202, the connecting block 203 is in an inverted V shape, a first clamping groove 204 and a second clamping groove 205 are formed in the connecting position of the connecting block 203 and the first separating cup 201 and the second separating cup 202 respectively, the first clamping groove 204 is clamped with the first guide plate 105, and the second clamping groove 205 is clamped with the second guide plate 106. In this embodiment, the first separating cup 201, the second separating cup 202 and the connecting block 203 are integrally formed, so that the integrally formed separating cup assembly 20 can reduce the processing cost and increase the structural stability and the sealing performance. The joint block 203 is provided with a limiting rod 206 in an outward protruding way, and the limiting hole 107 is matched with the limiting rod 206. The cooperation of the first clamping groove 204 and the first guide plate 105 and the cooperation of the second clamping groove 205 and the second guide plate 106 can enable the separation cup assembly 20 to limit in the length direction of the partition plate 101, and the cooperation of the limiting hole 107 and the limiting rod can enable the separation seat 10 and the separation cup assembly 20 not to be separated when being integrally inverted. Of course, in other embodiments, the separating cup assembly 20 and the separating base 10 may be detachably connected by a clamping or bonding method. It should be noted that the stop lever 206 may also be disposed on the first separation cup 201 and/or the second separation cup 202, and the stop holes 107 on the side plate 102 are in one-to-one correspondence with the stop levers 206. This three-point positioning structure of the present embodiment can smoothly fix the separation cup assembly 20 to the separation seat 10.

Equivalent embodiments of the above positioning structure will also be apparent to those skilled in the art, and they should fall within the scope of the present application. If a limit rod is arranged on the side plate 102, a limit hole is arranged on the connecting block 203.

In this embodiment, the magnet 30 is removable or insertable, and in other embodiments, the magnet 30 and the separating seat 10 may be integrally formed, so as to increase structural stability.

Preferably, the sides of the separating cup assembly 20 are planar, and the bottoms of the first separating cup 201 and the second separating cup 202 are planar to abut the partition 101. The flat-to-flat contact enables the separator cup assembly 20 to more smoothly ride on the separator plate 101.

In this embodiment, the separation cup assembly 20 is a transparent PC material that facilitates the observation of the cell mixture within the separation cups, preferably each separation cup having a capacity of 1mL.

Preferably, the upper end surfaces of the first guide plate 105 and the second guide plate 106 are rounded, and the first clamping groove 204 and the second clamping groove 205 are rounded. The clamping groove and the guide plate can be matched better through the round corner guiding treatment, and the processing difficulty is reduced.

It should be noted that in this embodiment, there are three separating cup assemblies 20, and six containers, and six cells can be separated at a time. In other embodiments, a plurality of sets of separating cup assemblies 20 may be provided by those skilled in the art, as the practice is not limited in this regard.

The cell separation device of the present embodiment has the following advantages:

1. The separating cup component is of a W-like shape, and the magnet can firmly adsorb the magnetic beads adsorbed with target cells at the bottom of the separating cup without wall hanging, so that the yield is improved;

2. The separating seat can be reused, the separating cup is disposable, and the cost is greatly reduced compared with the disposable separating column in the prior art;

3. Compared with the existing small magnetic bead separation technology, the separation procedure reduces the steps of washing a magnetic bead separation column, repeatedly washing for 3 times, washing target cells and centrifuging, reduces the steps of separating the magnetic bead cells, sucking the supernatant containing the target cells and centrifuging, shortens the reaction time, and reduces the cost of manpower, articles and time;

4. the whole process of cell separation is completed in one cup, so that cross contamination caused by tube replacement or container replacement in the separation process in the prior art is reduced;

5. The whole device has simple structure, convenient disassembly and assembly and easy cleaning and disinfection.

It should be noted that, in the above embodiment, two adjacent separation cups are combined to form a separation cup assembly, in other embodiments, three or four separation cups may be combined according to practical situations, and a plurality of guide plate assemblies are correspondingly arranged to perform clamping fit, so long as the separation cups are in a V-shaped arrangement, all the embodiments are within the scope of the present application. The case where three separate cups are combined to form a separate cup assembly is briefly described below.

The adjacent three separating cups are combined to form a separating cup assembly, and the separating cup assembly is integrally formed.

In one embodiment, the step of separating cells using the separation device described above is as follows:

incubating the mixed solution of the magnetic beads coupled with the antibodies corresponding to the target cells and the cell sample solution at the temperature of 2-8 ℃ for 10-40 minutes, such as incubating for 30 minutes on ice. Preferably, the incubation temperature is 3℃to 8℃and more preferably 4 ℃.

Adding the mixed solution of the incubated magnetic beads and the cell sample liquid into a separation cup, clamping the separation cup above a separation seat, standing for 1-3min at room temperature, and precipitating the magnetic beads coupled with the target cells at the bottom of the separation cup under the action of a magnetic field.

Spreading sterile gauze on the biosafety cabinet, integrally inverting the separation cup and the separation seat, and sucking the solution in the separation cup by utilizing the water absorption of the gauze. Compared with the method of sucking the supernatant by using a liquid-transfering device, the step sucks the supernatant in the separating cup as much as possible, and the purity of the target cells is improved.

The separation cup was removed and the magnetic bead-coupled positive cells precipitated in the separation cup were used directly for the next step.

By implementing the embodiment, compared with a method of separating by using a separation column, the separation process reduces the steps of washing, 3 times of repeated washing, washing target cells and centrifuging the small magnetic bead separation column. Compared with the large magnetic bead separation method, the method reduces the steps of magnetic bead shearing, absorption of supernatant containing target cells and centrifugation. Can shorten the separation time and reduce the cost of labor, articles and time. And the device is simple and easy to clean and sterilize. Meanwhile, the separating seat can be reused, and the device is more flexible.

To increase the cell purity, 50-500. Mu.L of cell culture medium may be added to the precipitated beads and the coupled cells, the beads and cells resuspended, and the above steps repeated. The washing step is completed. Washing can be selected for 1-3 times, and the washing step can improve the purity of the cells by 5%.

In another embodiment, the step of separating cells using the separation device described above is as follows:

Incubating the mixture of one or more magnetic beads coupled with the non-target cell-corresponding antibodies and the cell sample liquid at 2-8deg.C for 10-40 min, such as on ice for 30min. Preferably, the incubation temperature is 3℃to 8℃and more preferably 4 ℃.

Adding the mixed solution of the incubated magnetic beads and the cell sample liquid into a separation cup, clamping the separation cup above a separation seat, standing for 1-3min at room temperature, and precipitating the magnetic beads coupled with non-target cells to the bottom of the separation cup under the action of a magnetic field.

The cells in the supernatant in the separation cup are recovered and the non-target cells that have settled at the bottom of the separation cup due to the adsorption of the magnetic beads by the magnetic adsorption are discarded. The supernatant was carefully aspirated with a pipette without touching the beads and transferred to another separation cup (or sterile container). The cells contained in the supernatant are the target cells.

Similarly, in order to increase the cell purity, the washing step may be completed by repeating the above steps by adding again magnetic beads coupled with antibodies corresponding to non-target cells to the supernatant. Washing can be selected for 1-3 times.

Referring to fig. 4, fig. 4 is a schematic flow chart of a first embodiment of the cell separation method of the present application. In this embodiment, the cell separation method is performed by using the cell separation apparatus of any one of the above examples or embodiments, and comprises the steps of:

s410: providing a cell sample liquid, wherein the cell sample liquid at least comprises first-type cells.

The cell sample liquid may be obtained by processing an acquired biological sample, and different biological samples may be processed according to an existing corresponding processing method, which is not limited herein.

S420: the cell sample liquid and the first type magnetic beads are mixed in the separation cup, and the first type magnetic beads are coupled with the first type antibodies and are used for combining with the first type cells.

Wherein the diameter of the first type of magnetic beads is 500-5000 nanometers. Such as 500nm, 1000 nm, 1500 nm, 2000 nm, 4000 nm, etc. In the method, the magnetic beads have medium particle size, no damage to cells and high separation purity.

S430: the separation cup with the first type of magnetic beads and the cell sample liquid is placed over the separation seat with the magnet so that the magnet adsorbs the first type of magnetic beads to the bottom of the separation cup.

The first type of magnetic beads bind to the first cells using the first type of antibodies.

S440: separating the first type of magnetic beads from the cell supernatant.

Wherein, the magnetic beads are adsorbed by the magnetic field to separate the cells combined with the magnetic beads from other cells, and the cell supernatant can be removed by adopting a direct pouring mode.

By implementing this embodiment, when separating cells using small magnetic beads, the separation column is not used any more, but the magnetic beads and the cell supernatant are directly separated, so that the separation speed can be increased, the separation cost can be reduced, and the separation purity can be improved.

The first target cells are first type cells which are precipitated in a separation cup and are coupled with the first type magnetic beads, and the first type cells coupled with the first type magnetic beads can be directly used for the next operation. Specifically, the separating cup is removed from the separating seat, and the cell culture solution is added into the separating cup to obtain a first target cell sample solution which can be directly used for the next operation. Of course, the first type magnetic beads can be separated from the first type cells and then used for the next operation.

In one embodiment, the cell sample liquid may be added to the separation cup, and the first type magnetic beads may be added to the cell sample liquid in an amount of 0.3 to 2.0 times, and mixed in the separation cup to obtain the cell mixture liquid. The cell sample liquid and the first type magnetic beads of 0.3 to 2.0 times may be mixed to obtain a cell mixed liquid, and the cell mixed liquid may be added into a separation cup without limitation.

In one embodiment, after the first type magnetic beads are added into the cell sample liquid, the first type magnetic beads and the first type cells can be fully mixed and combined by using an oscillator or a mixer, so that the separation efficiency is improved. The shaking or homogenization speed should be controlled during the mixing process to ensure that the cells are not damaged. After the first type magnetic beads and the first type cells are fully mixed, a separation cup containing a cell mixed solution is placed on a separation seat.

In one embodiment, the first type of magnetic beads are added in an amount of 0.3 to 2.0 times the amount of the cell sample fluid, such as 0.3, 0.5, 1.0, 1.3, 1.8, 2.0, etc. The method has the advantages of small particle size of the magnetic beads and strong adsorption capacity, so that the amount of the magnetic beads is small, the use amount of the magnetic beads is reduced, and the cost is reduced.

In one embodiment, after removal of the cell supernatant, a cell culture solution may also be added to the first type of magnetic beads to wash the first type of cells. If the cell culture solution is added, the magnetic beads and the cells are resuspended, and then the magnetic field is put into the cell culture solution to adsorb and remove the clear liquid, thus completing the washing step. Can be washed for 1 to 3 times. The purity of the isolated cells can be increased by washing, which can increase the purity of the isolated cells by at least 5%.

After washing, adding a cell culture solution into the first type of magnetic beads; the beads and cells were resuspended to obtain a first type of cells. The magnetic beads are composed of nontoxic ferric oxide and polysaccharide, and can be biodegraded, so that the subsequent operation can be directly carried out without separation.

In one embodiment, the method can be used to isolate positive cells, where the magnetic beads used are those with antibodies corresponding to positive cells. Of course, also can be used to isolate negative cells. The magnetic beads can be combined with the target cells by adopting a direct magnetic cell labeling mode. Indirect magnetic cell labeling can also be adopted, and the magnetic beads are combined with unwanted cells, so that the target cells are in the cell supernatant, and the cell supernatant can be continuously sorted to obtain the target cells. The cells in the cell supernatant can be further sorted, specifically, the cell sample liquid further comprises a second type of cells, the cell supernatant and the second type of magnetic beads are mixed in another separation cup, and the second type of antibodies are coupled to the second type of magnetic beads and are used for being combined with the second type of cells; placing a separation cup with second type magnetic beads and cell supernatant above a separation seat with a magnet; separating the second type of magnetic beads from the cell supernatant; and removing the separating cup from the separating seat to obtain second target cells, wherein the second target cells are the second type of cells which are precipitated in the separating cup and are coupled with the second type of magnetic beads, and the obtained second target cells can be directly used for cell application and other operations.

The foregoing description is only of embodiments of the present application, and is not intended to limit the scope of the application, and all equivalent structures or equivalent processes using the descriptions and the drawings of the present application or directly or indirectly applied to other related technical fields are included in the scope of the present application.

Claims

1. The cell separation device is characterized by comprising a separation seat, at least one separation cup and at least one magnet, wherein the separation cup is used for containing a cell mixed solution with magnetic beads, the separation cup is detachably arranged on the separation seat, the magnet is arranged on the separation seat, and the magnet is arranged below the separation cup and is used for adsorbing the magnetic beads to the bottom of the separation cup; the vertical section of the separation cup is V-shaped, two adjacent separation cups are combined to form a separation cup assembly, the separation cup assembly is integrally formed, the separation cup assembly is W-shaped, the separation seat comprises a partition plate, a plurality of accommodating grooves are formed below the partition plate and used for accommodating the magnets, a guide plate assembly perpendicular to the partition plate is arranged above the partition plate, and the guide plate assembly comprises a first guide plate and a second guide plate which are arranged in parallel at intervals; the separating cup assembly comprises a first separating cup, a second separating cup and a connecting block for connecting the first separating cup and the second separating cup, wherein the connecting block is in an inverted V shape, a first clamping groove and a second clamping groove are formed in the connecting position of the connecting block and the first separating cup and the connecting position of the connecting block and the second separating cup respectively, the first clamping groove is clamped with the first guide plate, and the second clamping groove is clamped with the second guide plate; the separating seat further comprises a side plate, the side plate is arranged on one side of the partition plate and is perpendicular to the partition plate and the guide plate assembly, a limiting hole is formed in the side plate, a limiting rod is arranged on the connecting block in an outward protruding mode, and the limiting hole is matched with the limiting rod.

2. The cell separation apparatus according to claim 1, wherein the separation cup assemblies each have a flat side surface, and the bottoms of the first separation cup and the second separation cup are each flat to abut against the partition plate.

3. The cell separation apparatus according to claim 2, wherein the upper end surfaces of the first guide plate and the second guide plate are rounded, and the first clamping groove and the second clamping groove are rounded.

4. The cell separation apparatus according to claim 1, wherein the separation housing further comprises a packing plate parallel to the side plate for packing the magnet.