Apparatus and method for fully sealed and fully automated dispensing of cells

Abstract

An apparatus and method for fully sealed and fully automated cell dispensing. The apparatus includes a sample supply module, a liquid refill module, and a liquid distribution module. The apparatus allows cell dispensing to be performed in a sealed state, reducing the risk of infection due to contact with the external environment and ensuring the viability of dispensed cells, thereby increasing the accuracy of cells obtained after dispensing and improving the quality of dispensed cells.

Description

[Technical field]

[0001] The present invention relates to the field of biology, and in particular to an apparatus and method for fully sealed and fully automated dispensing of cells. [Background technology]

[0002] Cellular immunotherapy is a cell therapy to enhance the targeting ability, lethality and persistence of immune cells by collecting the patient's immune cells, genetic modification and in vitro amplification culture. In recent years, this treatment has had great clinical results in tumor immunotherapy, and it is expected to clinically cure tumors. However, the prepared immune cells need to be washed, concentrated and dispensed into multiple frozen bags, and the intermediate links such as the process flow of dispensing cells, equipment and facilities, and the selection of reagents will affect the quality of the dispensed cells, thereby affecting the clinical effect. Therefore, the process for fully sealed and fully automated dispensing of immune cells can ensure the safety and batch-to-batch stability of the product, reduce the effects caused by humans and the environment, and improve the quality of immune cell therapy products. Traditional cell dispensing can only be performed under artificial and non-sealed conditions, thereby increasing the risk of infection due to contact with the external environment and greatly affecting the safety of the product.

[0003] Therefore, there is a need to develop in the field a method for fully sealed and fully automated dispensing of immune cells, which can be used to rapidly obtain high quality immune cells for clinical use. Summary of the Invention [Problem to be solved by the invention]

[0004] It is an object of the present invention to provide a method for fully sealed and fully automated dispensing of immune cells that can be used to rapidly obtain high quality immune cells for clinical use. [Means for solving the problem]

[0005] In a first aspect, the present invention provides an apparatus for fully confined and fully automated dispensing of cells, the apparatus comprising a sample supply module, a liquid replenishment module and a liquid dispensing module, The sample supply module includes a cell sample container (such as a cell sample bag), The liquid refilling module includes a plurality of liquid refilling means, the liquid refilling means including a plurality of liquid refilling containers (e.g., liquid refilling bags (8-1, 8-2)); The liquid distribution module includes a liquid distributor, and the liquid dispenser is provided with a sample supply pipe, a liquid outlet pipe, an extraction pipe, a waste liquid pipe, and a liquid refill pipe connected thereto, the sample supply pipe, the liquid outlet pipe, the extraction pipe, the waste liquid pipe, and the liquid refill pipe are connected to each other, and the sample supply pipe, the liquid outlet pipe, the extraction pipe, the waste liquid pipe, and the liquid refill pipe are each provided with a valve; The sample supply pipe is connected to a cell sample container (such as a cell sample bag), the liquid outlet pipe is connected to a cell freezing container (such as a cell freezing bag (7)), the extraction pipe is connected to the extraction device (1), the waste pipe is connected to a waste liquid container (such as a waste liquid bag (6)), and the liquid replenishment pipe is connected in sequence to a number of liquid replenishment containers (such as liquid replenishment bags) via a number of branch pipes, and each branch pipe is provided with a valve.

[0006] In another preferred embodiment, the number of liquid refill containers (such as liquid refill bags) is 1, 2, 3, 4, 5, 6, 7, or 8.

[0007] In another preferred embodiment, the number of liquid replacement containers (such as liquid replacement bags) is two, and two different liquid replacement containers (such as liquid replacement bags) respectively contain Liquid Replacement Component 1 and Liquid Replacement Component 2. Preferably, Liquid Replacement Component 1 contains the compound electrolyte injection solution and the human serum albumin aqueous solution, and Liquid Replacement Component 2 contains the CS10 cryopreservation solution.

[0008] In another preferred embodiment, a cell sample container (such as a cell sample bag) contains the cell sample.

[0009] In another preferred embodiment, the compound electrolyte injection solution comprises 3 to 7 parts by weight of sodium chloride, 3 to 7 parts by weight of sodium gluconate, 2 to 6 parts by weight of sodium acetate, 0.1 to 0.8 parts by weight of potassium chloride, 0.1 to 0.6 parts by weight of magnesium chloride, and 950 to 1,050 parts by weight of water.

[0010] In another preferred embodiment, the concentration of human serum albumin in the aqueous human serum albumin solution is 15 to 25% (v / v), preferably 18 to 22% (v / v).

[0011] In another preferred embodiment, the volume ratio of the compound electrolyte injection solution to the aqueous human serum albumin solution is (80-120):1, preferably (90-110):1, and more preferably (95-105):1.

[0012] In another preferred embodiment, the CS10 cryopreservation medium comprises an aqueous DMSO solution.

[0013] In another preferred embodiment, the CS10 cryopreservation solution contains an 8 to 12% (v / v) aqueous DMSO solution.

[0014] In another preferred embodiment, the extractor is capable of aspirating liquid and pumping liquid.

[0015] In a second aspect, the present invention provides a method for preparing a cell suspension using the device according to the first aspect of the invention, the method comprising the steps of: (1) A cell sample is placed in a cell sample container (such as a cell sample bag), and a liquid supply component 1 and a liquid supply component 2 are placed in two different liquid supply containers (such as liquid supply bags), respectively; Fluid replacement component 1 comprises a compound electrolyte injection solution and a human serum albumin aqueous solution, and fluid replacement component 2 comprises a CS10 cryopreservation solution; (2) sending a cell sample in a cell sample container (e.g., a cell sample bag) into an extractor through a sample supply tube and an extraction tube based on suction of the extractor, and then sending the cell sample into a waste container (e.g., a waste bag) through an extraction tube and a waste tube based on discharge of the extractor, thereby performing tube washing of the sample supply tube, the extraction tube, and the waste tube; (3) sending a cell sample from a cell sample container (such as a cell sample bag) into the extractor through a sample supply tube and an extraction tube based on the suction of the extractor, and then sending the cell sample into a cell freezing container (such as a cell freezing bag) through the extraction tube and a liquid outlet tube based on the discharge of the extractor; (4) sending the liquid refill component 1 of the liquid refill container (such as a liquid refill bag) into the extractor through the liquid refill tube and the extraction tube based on the suction of the extractor, and then sending the liquid refill component 1 into the cell freezing container (such as a cell freezing bag) through the extraction tube and the liquid outlet tube based on the discharge of the extractor; and sending the liquid refill component 2 of the other liquid refill container (such as the other liquid refill bag) into the extractor through the liquid refill tube and the extraction tube based on the suction of the extractor, and then sending the liquid refill component 2 into the cell freezing container (such as the cell freezing bag) through the extraction tube and the liquid outlet tube based on the discharge of the extractor; In step (4), Liquid replenishment component 1 and Liquid replenishment component 2 are sequentially fed into a cell freezing container (eg, a cell freezing bag), and the volume ratio of Liquid replenishment component 1 to Liquid replenishment component 2 is 1:1.

[0016] In another preferred embodiment, the method further comprises step (5): counting the cells in the cell freezing container (such as a cell freezing bag) in step (4), performing liquid replenishment according to the operations of step (4) if the cell density is high, and replenishing the cell sample according to the operations of step (2) if the cell density is low, to obtain a cell suspension with a desired cell density.

[0017] In another preferred embodiment, the method further comprises a dispensing step (6), which comprises the steps of connecting a cell freezing container (such as a cell freezing bag) containing the cell suspension to a sample supply tube, and connecting a waste tube to a dispensing bag; The cell suspension in the cell freezing container (e.g., cell freezing bag) is sent into the extractor through a sample supply tube and an extraction tube based on the suction of the extractor, and then the cell suspension is sent into a dispensing bag through an extraction tube and a waste tube based on the discharge of the extractor, and multiple dispensing of the cell suspension is realized by changing the dispensing bag.

[0018] In another preferred embodiment, the dispensing step (6) is performed after step (5).

[0019] In another preferred embodiment, the compound electrolyte injection solution comprises 3 to 7 parts by weight of sodium chloride, 3 to 7 parts by weight of sodium gluconate, 2 to 6 parts by weight of sodium acetate, 0.1 to 0.8 parts by weight of potassium chloride, 0.1 to 0.6 parts by weight of magnesium chloride, and 950 to 1,050 parts by weight of water.

[0020] It should be understood that within the scope of the present invention, the above technical features of the present invention and the technical features specifically described below (such as examples) can be combined with each other to form a new or preferred technical scheme, which will not be described in detail in this specification due to space limitations.

[0021] The drawings are used only for illustrative description and should not be construed as limitations of the utility model. In order to better explain the embodiments, some parts of the drawings may be omitted, enlarged, or reduced, and do not represent the actual size of the product. It will be understood by those skilled in the art that some well-known structures and descriptions in the drawings may be omitted. The same or similar labels correspond to the same or similar parts. The terms used to describe the positional relationship in the drawings are used only for illustrative description and should not be construed as limitations of this patent. [Brief description of the drawings]

[0022] [Figure 1] FIG. 1 is a structural schematic diagram of an apparatus for complete sealing and fully automatic dispensing of cells in one example, in which 8-1 and 8-2 represent liquid refill bags, 5 represents a sample supply tube, 1 represents an extraction device, 6 represents a waste liquid bag, and 7 represents a cell freezing container (such as a cell freezing bag). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0023] Through extensive and detailed research, the present inventors have unexpectedly developed an apparatus and method for completely sealing and fully automatic dispensing of cells for the first time. This method allows cell dispensing to be performed under sealed conditions, reducing the risk of infection due to contact with the external environment. According to the apparatus and method for completely sealing and fully automatic dispensing of cells of the present invention, the survival rate of dispensed cells can be ensured, the accuracy of cells obtained after dispensing is high, and the quality of dispensed cells is improved. This has led the present inventors to complete the present invention.

[0024] term Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0025] As used herein, the terms "include," "comprise," and "contain" may be used interchangeably and include open definitions as well as semi-closed and closed definitions. In other words, the terms include "consisting of" and "consisting essentially of."

[0026] Device For ease of explanation, the device for fully sealed and fully automated dispensing of cells of the present invention is further described below with reference to Figure 1. It should be understood that the drawings in this invention are not intended to limit the scope of the invention.

[0027] Typically, the device for fully sealing and fully automatic dispensing of cells in the present invention includes a sample supply module, a liquid replenishment module, and a liquid distribution module; the sample supply module includes a cell sample container (such as a cell sample bag); The liquid refill module includes a plurality of liquid refill means, the liquid refill means including a plurality of liquid refill containers (e.g., liquid refill bags (8-1, 8-2)); The liquid distribution module includes a liquid distributor, and the liquid dispenser is provided with a sample supply pipe, a liquid outlet pipe, an extraction pipe, a waste liquid pipe, and a liquid refill pipe connected thereto, the sample supply pipe, the liquid outlet pipe, the extraction pipe, the waste liquid pipe, and the liquid refill pipe are connected to each other, and the sample supply pipe, the liquid outlet pipe, the extraction pipe, the waste liquid pipe, and the liquid refill pipe are each provided with a valve; The sample supply pipe is connected to a cell sample container (such as a cell sample bag), the liquid outlet pipe is connected to a cell freezing container (such as a cell freezing bag (7)), the extraction pipe is connected to the extraction device (1), the waste pipe is connected to a waste liquid container (such as a waste liquid bag (6)), and the liquid replenishment pipe is connected in sequence to a number of liquid replenishment containers (such as liquid replenishment bags) via a number of branch pipes, and each branch pipe is provided with a valve.

[0028] In a preferred embodiment of the present invention, the number of liquid refill containers (such as liquid refill bags) is 1, 2, 3, 4, 5, 6, 7, or 8.

[0029] In another preferred embodiment of the present invention, the number of liquid replacement containers (such as liquid replacement bags) is two, and two different liquid replacement containers (such as liquid replacement bags) respectively contain liquid replacement component 1 and liquid replacement component 2. Preferably, liquid replacement component 1 contains a compound electrolyte injection solution and a human serum albumin aqueous solution, and liquid replacement component 2 contains a CS10 cryopreservation solution.

[0030] Preferably, the compound electrolyte injection solution contains 3 to 7 parts by weight of sodium chloride, 3 to 7 parts by weight of sodium gluconate, 2 to 6 parts by weight of sodium acetate, 0.1 to 0.8 parts by weight of potassium chloride, 0.1 to 0.6 parts by weight of magnesium chloride, and 950 to 1,050 parts by weight of water.

[0031] In another preferred embodiment, the concentration of human serum albumin in the aqueous human serum albumin solution is 15 to 25% (v / v), preferably 18 to 22% (v / v).

[0032] In another preferred embodiment, the volume ratio of the compound electrolyte injection solution to the aqueous human serum albumin solution is (80-120):1, preferably (90-110):1, and more preferably (95-105):1.

[0033] In another preferred embodiment, the CS10 cryopreservation medium comprises an aqueous DMSO solution.

[0034] In another preferred embodiment, the CS10 cryopreservation solution contains an 8 to 12% (v / v) aqueous DMSO solution.

[0035] In another preferred embodiment of the invention, a cell sample container (such as a cell sample bag) contains the cell sample.

[0036] method The present invention further provides a method for dispensing cells using the device for fully sealed and fully automated dispensing of cells of the present invention, the method comprising the steps of: (1) A cell sample is placed in a cell sample container (such as a cell sample bag), and a liquid supply component 1 and a liquid supply component 2 are placed in two different liquid supply containers (such as liquid supply bags), respectively; Fluid replacement component 1 comprises a compound electrolyte injection solution and a human serum albumin aqueous solution, and fluid replacement component 2 comprises a CS10 cryopreservation solution; (2) sending a cell sample in a cell sample container (e.g., a cell sample bag) into an extractor through a sample supply tube and an extraction tube based on suction of the extractor, and then sending the cell sample into a waste container (e.g., a waste bag) through an extraction tube and a waste tube based on discharge of the extractor, thereby performing tube washing of the sample supply tube, the extraction tube, and the waste tube; (3) sending a cell sample from a cell sample container (such as a cell sample bag) into the extractor through a sample supply tube and an extraction tube based on the suction of the extractor, and then sending the cell sample into a cell freezing container (such as a cell freezing bag) through the extraction tube and a liquid outlet tube based on the discharge of the extractor; (4) sending the liquid refill component 1 of the liquid refill container (such as a liquid refill bag) into the extractor through the liquid refill tube and the extraction tube based on the suction of the extractor, and then sending the liquid refill component 1 into the cell freezing container (such as a cell freezing bag) through the extraction tube and the liquid outlet tube based on the discharge of the extractor; and sending the liquid refill component 2 of the other liquid refill container (such as the other liquid refill bag) into the extractor through the liquid refill tube and the extraction tube based on the suction of the extractor, and then sending the liquid refill component 2 into the cell freezing container (such as the cell freezing bag) through the extraction tube and the liquid outlet tube based on the discharge of the extractor; In step (4), Liquid replenishment component 1 and Liquid replenishment component 2 are sequentially fed into a cell freezing container (eg, a cell freezing bag), and the volume ratio of Liquid replenishment component 1 to Liquid replenishment component 2 is 1:1.

[0037] In a preferred embodiment of the present invention, the compound electrolyte injection solution contains 3 to 7 parts by weight of sodium chloride, 3 to 7 parts by weight of sodium gluconate, 2 to 6 parts by weight of sodium acetate, 0.1 to 0.8 parts by weight of potassium chloride, 0.1 to 0.6 parts by weight of magnesium chloride, and 950 to 1,050 parts by weight of water.

[0038] In a preferred embodiment of the present invention, the method further includes step (5): counting the cells in the cell freezing container (such as a cell freezing bag) in step (4), performing liquid replenishment according to the operation of step (4) if the cell density is high, and replenishing the cell sample according to the operation of step (2) if the cell density is low, to obtain a cell suspension with a desired cell density.

[0039] In a preferred embodiment of the present invention, the method further comprises a dispensing step (6), which comprises the steps of connecting a cell freezing container (such as a cell freezing bag) containing a cell suspension to a sample supply tube, and connecting a waste tube to a dispensing bag; The cell suspension in the cell freezing container (e.g., cell freezing bag) is sent into the extractor through a sample supply tube and an extraction tube based on the suction of the extractor, and then the cell suspension is sent into a dispensing bag through an extraction tube and a waste tube based on the discharge of the extractor, and multiple dispensing of the cell suspension is realized by changing the dispensing bag.

[0040] In another preferred embodiment, the dispensing step (6) is performed after step (5).

[0041] The present invention has the following main advantages:

[0042] According to the device and method for completely sealing and fully automatic dispensing of cells of the present invention, cell dispensing can be performed under sealed conditions, thereby reducing the risk of infection due to contact with the external environment. According to the device and method for completely sealing and fully automatic dispensing of cells of the present invention, the survival rate of dispensed cells can be ensured, the accuracy of cells obtained after dispensing is high, and the quality of dispensed cells is improved.

[0043] The method for preparing cells of the present invention can rapidly prepare immune cells, reduce the cost of enterprises, improve production capacity, and be suitable for industrial production. Meanwhile, the immune cells prepared by the method for preparing cells of the present invention have high quality and can ensure clinical efficacy.

[0044] The present invention will be further described below in combination with specific examples.It is understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.The experimental methods without specific conditions in the following examples are usually used according to conventional conditions or according to the conditions recommended by manufacturers.Unless otherwise stated, percentages and parts are calculated by weight.

[0045] [Example 1] 1. Experimental Scheme a) Number of cryopreserved cells required: The amount of positive T cells in the final product bag is 150 x 10 6 The cell was frozen and the volume of frozen storage was 40 ml, and the amount of positive T cells in the test cell bag was 37.5 × 10 6 The cryopreservation volume is 10 ml, the cell positive rate is 20.71%, and the theoretical cryopreservation density is 19.918 × 10 6 cells / ml.

[0046] b) Preparation of functional reagents: i. Component 1: HSA (human serum albumin) aqueous solution with a concentration of 20% (v / v) was added to compound electrolyte injection solution (containing sodium chloride 5.26 g, sodium gluconate 5.02 g, sodium acetate 3.68 g, potassium chloride 0.37 g, and magnesium chloride 0.30 g, each in 1,000 ml water), and then filled into component 1 bag, and the volume ratio of HSA aqueous solution with a concentration of 20% (v / v) to compound electrolyte injection solution was 1:100.

[0047] ii. Component 2: CS10 cryopreservation solution was filled into the bags of component 2, CS10 cryopreservation solution was a DMSO aqueous solution, and the volume fraction of DMSO was 10%.

[0048] c) Preparation of pipes before dispensing: i. Kit CT-49.1 (Apparatus for Fully Containing and Fully Automatic Dispensing of Cells) was opened in a safety cabinet, the clamps were closed, and sterile tubing connections were used to connect the harvested cell sample bag at position 5, the 250 ml cell freezer bag at position 7, the bag of component 1 at position 8-1, the bag of component 2 at position 8-2, and the waste bag at position 6. A schematic diagram of the piping positions for Kit CT-49.1 is shown in Figure 1.

[0049] ii. The tube is installed in the Sepax according to Figure 1.

[0050] d) Concentration of cell suspension by culture washing procedure i. Cell counting: After concentrating the cell suspension, a 5 ml syringe was connected to the 250 ml freezer bag containing the cell suspension, the freezer bag was shaken evenly, and about 0.2 ml of the cell suspension was aspirated to perform cell counting.

[0051] e) Preparation of final product by dilution (aliquoting) procedure: Component 1 and Component 2 were added in a 50%:50% ratio to the final cryopreservation solution.

[0052] i. Component volume calculation: Based on the cell counting results in the above steps, the cell suspension volume, the final product packing density and the final product packing cell amount, the adding volumes of component 1 and component 2 were calculated respectively. Adding volume of component 1 = the volume of cell suspension in the freezer bag + the volume of component 1 that needs to be added further.

[0053] ii. Vascular rinse: The rinse volume of component 1, 10 ml, was imputed, the tubing clamp was opened, and the procedure was started.

[0054] iii. Dispensing component 1: attribute the dispensed volume, i.e. the volume of component 1 that still needs to be added, click "Re", open the tubing clamps and start the procedure.

[0055] iv. Vascular rinse: The rinse volume of component 2, 10 ml, was imputed, the tubing clamp was opened, and the procedure was started.

[0056] v. Dispense component 2: impute the dispense volume, i.e. the total volume of component 2 that needs to be added, click "Re", open the tubing clamps and start the procedure.

[0057] f) Dispensing of the final product: i. Vascular rinse: The final product rinse volume, 10 ml, was imputed, the tubing clamp was opened, and the procedure was started.

[0058] ii. Dispensing the test cell bag: Before dispensing, first calculate the dispensing volume according to the filled cell volume and cryopreservation density of the test cell bag, then connect the 50ml cell freezing bag to position 6, impute the calculated dispensing volume, click "Re", open the tube clamp, and start the procedure. This step was repeated twice to obtain three test cell bags.

[0059] iii. Final product dispensing: Before dispensing, first calculate the dispensing volume according to the filled cell volume and cryopreservation density of the final product bag, then connect the 250ml cell freezing bag to position 6, impute the calculated dispensing volume, click "Re", open the tubing clamp, and start the procedure. Finally, one final product bag was obtained.

[0060] iv. The air was evacuated from the three test cell bags and the final product bag and the bags were sealed.

[0061] v. The sealed bag was transferred to a constant speed freezer, frozen in the constant speed freezer, and then transferred to a liquid nitrogen freezer for freezing and storage.

[0062] 2. Experimental Results a) Counting of concentrated cell suspensions The density and viability of the concentrated cells in the cell sample bag were measured by a cell counter as shown in Table 1. The cell amount after concentration was 181.5 × 10 6 cells / ml×(12.2ml-0.2ml)=2178×10 6 It is written as cells. [Table 1]

[0063] b) Preparation of the final product The calculated component volumes are shown in Table 2, and the total volume is expressed as 54.7 ml + 54.7 ml = 109.4 ml. [Table 2]

[0064] c) Dispensing of the final product The density and viability of the cells dispensed into the three test cell bags and the final product bag were measured by a cell counter as shown in Tables 4, 5, 6 and 7, respectively. The amount of positive cells in test cell bag 1 was 19.25×10 6 cells / ml×10ml×20.71%=39.87×10 6 The amount of positive cells in test cell bag 2 was 18.8 × 10 6 cells / ml×10ml×20.71%=38.93×10 6 The amount of positive cells in test cell bag 3 was 16.5 × 10 6 cells / ml×10ml×20.71%=34.17×10 6 The amount of positive cells in the final product bag was 20.033 × 10 6 cells / ml×40ml×20.71%=165.95×10 6 The results were expressed as cells. The summary results of cell dispensing are shown in Table 8. Compared with the theoretical freezing density, there is no significant difference in the cell densities of the three test cell bags and the final product bag, with the maximum difference being 1517.16%. Compared with the theoretical positive cell volume, the maximum difference is 10.63%, which is within the acceptable range, and the various bags have little difference in cell viability. Therefore, the volume and total cell volume of the final product prepared by Sepax using this process are accurate, there is almost no loss of cells, and it is possible to perform final product dispensing using Sepax for the recovered cells, indicating that the fully sealed automation of the process flow of dispensing immune cells is realized. [Table 3] [Table 4] [Table 5] [Table 6] [Table 7]

[0065] All documents mentioned in the present invention are incorporated by reference in this application as if each document was incorporated by reference separately. In addition, after reading the above content of the present invention, those skilled in the art may make various changes or modifications to the present invention, and all equivalent forms are within the scope defined by the claims of this application.

Claims

1. An apparatus for automatic dispensing of cells, comprising a sample supply module, a liquid replenishment module, and a liquid dispensing module, wherein the sample supply module comprises a cell sample container, the liquid replenishment module comprises a plurality of liquid replenishment means, and the plurality of liquid replenishment means comprise a plurality of liquid replenishment containers, the liquid dispensing module comprises a liquid dispenser, and the liquid dispenser is provided with a connected sample supply pipe, a liquid outlet pipe, an extraction pipe, a waste liquid pipe, and a liquid replenishment pipe. The sample supply pipe, the liquid outlet pipe, the extraction pipe, the waste liquid pipe, and the liquid replenishment pipe communicate with each other, and valves are respectively provided on the sample supply pipe, the liquid outlet pipe, the extraction pipe, the waste liquid pipe, and the liquid replenishment pipe, the sample supply pipe is connected to the cell sample container, the liquid outlet pipe is connected to a cell freezing container, the extraction pipe is connected to an extraction device, the waste liquid pipe is connected to a waste liquid container, and the liquid replenishment pipe is sequentially connected to the plurality of liquid replenishment containers via a plurality of branch pipes, and each branch pipe is provided with a valve.

2. The number of the liquid replenishment containers is two, and the two different liquid replenishment containers respectively contain a liquid replenishment component 1 and a liquid replenishment component 2. The liquid replenishment component 1 contains a compound electrolyte injection solution and an aqueous human serum albumin solution, and the liquid replenishment component 2 contains a CS10 cryopreservation solution. The apparatus according to claim 1.

3. The compound electrolyte injection solution contains 3 to 7 parts by weight of sodium chloride, 3 to 7 parts by weight of sodium gluconate, 2 to 6 parts by weight of sodium acetate, 0.1 to 0.8 parts by weight of potassium chloride, 0.1 to 0.6 parts by weight of magnesium chloride, and 950 to 1,050 parts by weight of water. The apparatus according to claim 2.

4. The aqueous human serum albumin solution contains human serum albumin at a concentration of 15 to 25% (v / v). The apparatus according to claim 2.

5. The extraction device can suck and pump liquid. The apparatus according to claim 1.

6. The volume ratio of the compound electrolyte injection solution to the aqueous human serum albumin solution is (80 to 120):1, preferably (90 to 110):1, more preferably (95 to 105):

1. The apparatus according to claim 2.

7. The CS10 cryopreservation solution contains 8 to 12% (v / v) DMSO. The apparatus according to claim 2.

8. A method for preparing a cell suspension using the apparatus according to claim 1, comprising the following steps, namely, (1) adding a cell sample to the cell sample container, and adding liquid supplement component 1 and liquid supplement component 2 to two different liquid supplement containers respectively, wherein the liquid supplement component 1 contains a compound electrolyte injection solution and an aqueous human serum albumin solution, and the liquid supplement component 2 contains a CS10 cryopreservation solution; (2) sending the cell sample in the cell sample container into the extraction device through the sample supply pipe and the extraction pipe, and then sending the cell sample into the waste liquid container through the extraction pipe and the waste liquid pipe to perform pipe cleaning of the sample supply pipe, the extraction pipe and the waste liquid pipe; (3) sending the cell sample in the cell sample container into the extraction device through the sample supply pipe and the extraction pipe, and then sending the cell sample into the cell freezing container through the extraction pipe and the liquid outlet pipe; (4) sending the liquid supplement component 1 in the liquid supplement container into the extraction device through the liquid supplement pipe and the extraction pipe, and then sending the liquid supplement component 1 into the cell freezing container through the extraction pipe and the liquid outlet pipe; sending the liquid supplement component 2 in the other liquid supplement container into the extraction device through the liquid supplement pipe and the extraction pipe, and then sending the liquid supplement component 2 into the cell freezing container through the extraction pipe and the liquid outlet pipe; and in step (4), the liquid supplement component 1 and the liquid supplement component 2 are sequentially sent into the cell freezing container, and the volume ratio of the liquid supplement component 1 to the liquid supplement component 2 is 1:

1. Method.

9. Step (5): For obtaining a cell suspension with a desired cell density, counting the cells in the cell freezing container in step (4), and when the cell density is high, performing liquid supplementation according to step (4), and when the cell density is low, supplementing the cell sample according to step (2), further comprising the method according to claim 8.

10. Step (6) further comprising connecting the cell freezing container containing the cell suspension to the sample supply pipe and connecting the waste liquid pipe to a dispensing bag. The cell suspension in the cell freezing container is sent into the extraction device by the sample supply pipe and the extraction pipe, and then the cell suspension is sent into the dispensing bag by the extraction pipe and the waste liquid pipe to dispense the cell suspension. The method according to claim 8.

11. The compound electrolyte injection solution contains 3 to 7 parts by weight of sodium chloride, 3 to 7 parts by weight of sodium gluconate, 2 to 6 parts by weight of sodium acetate, 0.1 to 0.8 parts by weight of potassium chloride, 0.1 to 0.6 parts by weight of magnesium chloride, and 950 to 1,050 parts by weight of water. The method according to claim 8.

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