WO2023025192A1 - 一种全封闭和全自动化的分装细胞的装置和方法 - Google Patents
一种全封闭和全自动化的分装细胞的装置和方法 Download PDFInfo
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- WO2023025192A1 WO2023025192A1 PCT/CN2022/114503 CN2022114503W WO2023025192A1 WO 2023025192 A1 WO2023025192 A1 WO 2023025192A1 CN 2022114503 W CN2022114503 W CN 2022114503W WO 2023025192 A1 WO2023025192 A1 WO 2023025192A1
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- tube
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- 238000000034 method Methods 0.000 title claims abstract description 30
- 239000007788 liquid Substances 0.000 claims abstract description 98
- 238000009826 distribution Methods 0.000 claims abstract description 7
- 238000000605 extraction Methods 0.000 claims description 90
- 238000007710 freezing Methods 0.000 claims description 46
- 230000008014 freezing Effects 0.000 claims description 46
- 239000002699 waste material Substances 0.000 claims description 36
- 238000002347 injection Methods 0.000 claims description 28
- 239000007924 injection Substances 0.000 claims description 28
- 239000006285 cell suspension Substances 0.000 claims description 25
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 21
- 239000007864 aqueous solution Substances 0.000 claims description 20
- 102000008100 Human Serum Albumin Human genes 0.000 claims description 18
- 108091006905 Human Serum Albumin Proteins 0.000 claims description 18
- 150000001875 compounds Chemical class 0.000 claims description 17
- 239000003792 electrolyte Substances 0.000 claims description 17
- 238000002637 fluid replacement therapy Methods 0.000 claims description 15
- 238000005070 sampling Methods 0.000 claims description 15
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- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 claims description 14
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 14
- 238000005138 cryopreservation Methods 0.000 claims description 14
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 12
- 238000003860 storage Methods 0.000 claims description 12
- AEQDJSLRWYMAQI-UHFFFAOYSA-N 2,3,9,10-tetramethoxy-6,8,13,13a-tetrahydro-5H-isoquinolino[2,1-b]isoquinoline Chemical compound C1CN2CC(C(=C(OC)C=C3)OC)=C3CC2C2=C1C=C(OC)C(OC)=C2 AEQDJSLRWYMAQI-UHFFFAOYSA-N 0.000 claims description 7
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 7
- 229910001629 magnesium chloride Inorganic materials 0.000 claims description 7
- 239000001632 sodium acetate Substances 0.000 claims description 7
- 235000017281 sodium acetate Nutrition 0.000 claims description 7
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- 235000011164 potassium chloride Nutrition 0.000 claims description 6
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- 210000004027 cell Anatomy 0.000 description 163
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Images
Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0236—Mechanical aspects
- A01N1/0242—Apparatuses, i.e. devices used in the process of preservation of living parts, such as pumps, refrigeration devices or any other devices featuring moving parts and/or temperature controlling components
- A01N1/0252—Temperature controlling refrigerating apparatus, i.e. devices used to actively control the temperature of a designated internal volume, e.g. refrigerators, freeze-drying apparatus or liquid nitrogen baths
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61J—CONTAINERS SPECIALLY ADAPTED FOR MEDICAL OR PHARMACEUTICAL PURPOSES; DEVICES OR METHODS SPECIALLY ADAPTED FOR BRINGING PHARMACEUTICAL PRODUCTS INTO PARTICULAR PHYSICAL OR ADMINISTERING FORMS; DEVICES FOR ADMINISTERING FOOD OR MEDICINES ORALLY; BABY COMFORTERS; DEVICES FOR RECEIVING SPITTLE
- A61J1/00—Containers specially adapted for medical or pharmaceutical purposes
- A61J1/14—Details; Accessories therefor
- A61J1/20—Arrangements for transferring or mixing fluids, e.g. from vial to syringe
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65B—MACHINES, APPARATUS OR DEVICES FOR, OR METHODS OF, PACKAGING ARTICLES OR MATERIALS; UNPACKING
- B65B3/00—Packaging plastic material, semiliquids, liquids or mixed solids and liquids, in individual containers or receptacles, e.g. bags, sacks, boxes, cartons, cans, or jars
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65B—MACHINES, APPARATUS OR DEVICES FOR, OR METHODS OF, PACKAGING ARTICLES OR MATERIALS; UNPACKING
- B65B3/00—Packaging plastic material, semiliquids, liquids or mixed solids and liquids, in individual containers or receptacles, e.g. bags, sacks, boxes, cartons, cans, or jars
- B65B3/04—Methods of, or means for, filling the material into the containers or receptacles
- B65B3/10—Methods of, or means for, filling the material into the containers or receptacles by application of pressure to material
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65B—MACHINES, APPARATUS OR DEVICES FOR, OR METHODS OF, PACKAGING ARTICLES OR MATERIALS; UNPACKING
- B65B3/00—Packaging plastic material, semiliquids, liquids or mixed solids and liquids, in individual containers or receptacles, e.g. bags, sacks, boxes, cartons, cans, or jars
- B65B3/26—Methods or devices for controlling the quantity of the material fed or filled
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65B—MACHINES, APPARATUS OR DEVICES FOR, OR METHODS OF, PACKAGING ARTICLES OR MATERIALS; UNPACKING
- B65B63/00—Auxiliary devices, not otherwise provided for, for operating on articles or materials to be packaged
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65B—MACHINES, APPARATUS OR DEVICES FOR, OR METHODS OF, PACKAGING ARTICLES OR MATERIALS; UNPACKING
- B65B65/00—Details peculiar to packaging machines and not otherwise provided for; Arrangements of such details
Definitions
- the invention relates to the field of biotechnology, in particular to a fully enclosed and fully automated cell subpackaging device and method.
- Cellular immunotherapy is a kind of cell therapy that collects the patient's own immune cells, undergoes genetic modification and in vitro expansion and culture, and then enhances the targeting, lethality and persistence of immune cells.
- tumor immunotherapy has performed very well clinically, bringing hope to the clinical cure of tumors.
- the prepared immune cells need to be washed, concentrated, and distributed into multiple cryopreservation bags. Any intermediate link such as the process flow, equipment, facilities, and reagent selection of the cells will affect the quality of the cells, and then affect the clinical effect. . Therefore, a fully enclosed and fully automated immune cell packaging process can ensure product safety and batch-to-batch stability, reduce the impact of personnel and the environment, and improve the quality of immune cell therapy products.
- Traditional cell subpackaging can only be carried out under artificial and non-closed conditions, which increases the risk of infection from contact with the external environment and greatly affects the safety of the product.
- the purpose of the present invention is to provide a fully enclosed and automatic method for subpackaging immune cells, which can quickly obtain high-quality immune cells for clinical use.
- a fully enclosed and fully automated device for distributing cells includes a sampling module, a liquid replenishment module and a liquid distribution module;
- the sample injection module includes a cell sample container (such as a cell sample bag);
- the fluid replacement module includes multiple fluid replacement devices, and the fluid replacement device includes multiple fluid replacement containers (such as fluid replacement bags (8-1, 8-2));
- the liquid distribution module includes a liquid distributor, and the liquid distributor is provided with a sample inlet tube, a liquid outlet tube, an extraction tube, a waste liquid tube and a liquid replacement tube, and the sample inlet tube, the liquid outlet tube, the extraction tube
- the pipe, the waste liquid pipe and the liquid replenishment pipe are connected, and valves are provided on the sampling pipe, liquid outlet pipe, extraction pipe, waste liquid pipe and liquid replenishment pipe;
- the sample inlet tube is connected to the cell sample container (such as a cell sample bag), the liquid outlet pipe is connected to the cell freezing container (such as a cell freezing bag (7), and the extraction tube is connected to the extraction device (1) connected, the waste liquid pipe is connected with the waste liquid container (such as waste liquid bag (6)), and the described liquid replenishment pipe is connected with a plurality of liquid replenishment containers (such as liquid replenishment bag) successively through a plurality of branch pipes, and each A valve is arranged on the branch pipe.
- the cell sample container such as a cell sample bag
- the liquid outlet pipe is connected to the cell freezing container (such as a cell freezing bag (7)
- the extraction tube is connected to the extraction device (1) connected
- the waste liquid pipe is connected with the waste liquid container (such as waste liquid bag (6))
- the described liquid replenishment pipe is connected with a plurality of liquid replenishment containers (such as liquid replenishment bag) successively through a plurality of branch pipes, and each A valve is arranged on the branch pipe.
- the number of the rehydration container (eg, rehydration bag) is 1, 2, 3, 4, 5, 6, 7 or 8.
- rehydration containers such as rehydration bags
- two different rehydration containers such as rehydration bags
- rehydration component 1 and rehydration component 2 respectively.
- the rehydration component 1 includes compound electrolyte injection and human serum albumin aqueous solution
- the rehydration component 2 includes CS10 frozen storage solution.
- the cell sample container (such as a cell sample bag) contains cell samples.
- the compound electrolyte injection includes 3-7 parts by weight of sodium chloride, 3-7 parts by weight of sodium gluconate, 2-6 parts by weight of sodium acetate, 0.1-0.8 parts by weight of potassium chloride, 0.1-0.6 parts by weight of magnesium chloride and 950-1050 parts by weight of water.
- the concentration of human serum albumin in the aqueous solution of human serum albumin is 15-25% (v/v), preferably 18-22% (v/v).
- the volume ratio of the compound electrolyte injection to the human serum albumin aqueous solution is 80-120:1, preferably 90-110:1, more preferably 95-105:1.
- the CS10 cryopreservation solution includes DMSO aqueous solution.
- the CS10 cryopreservation solution includes 8-12% (v/v) DMSO aqueous solution.
- the pumping device is capable of sucking in liquid and pushing out liquid.
- a method for preparing a cell suspension using the device as described in the first aspect of the present invention comprising the steps of:
- the cell sample container (such as a cell sample bag) contains a cell sample
- two different rehydration containers (such as a rehydration bag) respectively contain a rehydration component 1 and a rehydration component 2, wherein,
- the rehydration component 1 includes compound electrolyte injection and human serum albumin aqueous solution, and the rehydration component 2 includes CS10 frozen storage solution;
- the cell sample in the cell sample container (such as a cell sample bag) enters the extraction device through the injection tube and the extraction tube, and then the cell sample is discharged through the extraction tube and the extraction device.
- the waste liquid tube enters into the waste liquid container (such as waste liquid bag), so as to perform pipeline rinsing on the sampling tube, extraction tube and waste liquid tube;
- the cell sample in the cell sample container (such as a cell sample bag) enters the extraction device through the injection tube and the extraction tube, and then the cell sample is discharged through the extraction tube and The liquid outlet pipe enters into the cell freezing container (such as a cell freezing bag);
- the rehydration component 1 in the rehydration container (such as a rehydration bag) enters the withdrawal device through the rehydration tube and the extraction tube, and then the rehydration component 1 passes through the extraction tube and the extraction tube through the discharge of the extraction device.
- the liquid outlet pipe enters into the cell freezing container (such as a cell freezing bag);
- the rehydration component 2 in the rehydration container (such as a rehydration bag) enters the withdrawal device through the rehydration tube and the extraction tube, and then through the discharge of the extraction device, the rehydration component 2 passes through the extraction tube and the liquid outlet tube into the cell freezing container (such as a cell freezing bag);
- step (4) the rehydration component 1 and the rehydration component 2 enter into the cell freezing container (such as a cell freezing bag) sequentially, and the volume ratio of the rehydration component 1 and the rehydration component 2 is 1:1.
- the cell freezing container such as a cell freezing bag
- the method further includes step (5): counting the cells in the cell freezing container (such as a cell freezing bag) described in step (4), if the cell density is relatively large, follow the above steps ( Step 4) for rehydration, if the cell density is small, follow the steps of the above step (2) to replenish the cell sample, so as to obtain the cell suspension with the required cell density.
- the cell freezing container such as a cell freezing bag
- the method further includes the subpackaging step (6), which includes: connecting the cell freezing container (such as a cell freezing bag) containing the cell suspension to the sampling tube, The waste liquid pipe is connected with the sub-packaging bag;
- the cell suspension in the cell freezing container (such as a cell freezing bag) enters the extraction device through the injection tube and the extraction tube, and then the cell suspension passes through the extraction tube and the waste through the discharge of the extraction device.
- the liquid pipe enters into the subpackaging bag, and multiple subpackaging of the cell suspension is realized by changing the subpackaging bag.
- the subpackaging step (6) is after the step (5).
- the compound electrolyte injection includes 3-7 parts by weight of sodium chloride, 3-7 parts by weight of sodium gluconate, 2-6 parts by weight of sodium acetate, 0.1-0.8 parts by weight of potassium chloride, 0.1-0.6 parts by weight of magnesium chloride and 950-1050 parts by weight of water.
- Fig. 1 is a structural schematic diagram of an embodiment of a fully enclosed and fully automated device for subpackaging cells, wherein 8-1 and 8-2 are liquid replacement bags, 5 is a sampling tube, 1 is a extraction device, 6 is a waste liquid bag and 7 is a cell freezing container (such as a cell freezing bag).
- 8-1 and 8-2 are liquid replacement bags
- 5 is a sampling tube
- 1 is a extraction device
- 6 is a waste liquid bag
- 7 is a cell freezing container (such as a cell freezing bag).
- the inventor accidentally developed a fully enclosed and fully automated device and method for subpackaging cells for the first time.
- the method can perform cell subpackaging under closed conditions and reduce contact with the external environment.
- the risk of infection, the fully enclosed and fully automated device and method for subpackaging cells described in the present invention can ensure the survival rate of subpackaged cells, and the precision of the subpackaged cells is high, thereby improving the quality of the subpackaged cells.
- the inventors have completed the present invention.
- the terms “comprising”, “comprising” and “containing” are used interchangeably to include not only open definitions, but also semi-closed, and closed definitions. In other words, the terms include “consisting of”, “consisting essentially of”.
- the fully enclosed and fully automated cell subpackaging device of the present invention includes a sampling module, a liquid replenishment module and a liquid distribution module;
- the sample injection module includes a cell sample container (such as a cell sample bag);
- the fluid replacement module includes multiple fluid replacement devices, and the fluid replacement device includes multiple fluid replacement containers (such as fluid replacement bags (8-1, 8-2));
- the liquid distribution module includes a liquid distributor, and the liquid distributor is provided with a sample inlet tube, a liquid outlet tube, an extraction tube, a waste liquid tube and a liquid replacement tube, and the sample inlet tube, the liquid outlet tube, the extraction tube
- the pipe, the waste liquid pipe and the liquid replenishment pipe are connected, and valves are provided on the sampling pipe, liquid outlet pipe, extraction pipe, waste liquid pipe and liquid replenishment pipe;
- the sample inlet tube is connected to the cell sample container (such as a cell sample bag), the liquid outlet pipe is connected to the cell freezing container (such as a cell freezing bag (7), and the extraction tube is connected to the extraction device (1) connected, the waste liquid pipe is connected with the waste liquid container (such as waste liquid bag (6)), and the described liquid replenishment pipe is connected with a plurality of liquid replenishment containers (such as liquid replenishment bag) successively through a plurality of branch pipes, and each A valve is arranged on the branch pipe.
- the cell sample container such as a cell sample bag
- the liquid outlet pipe is connected to the cell freezing container (such as a cell freezing bag (7)
- the extraction tube is connected to the extraction device (1) connected
- the waste liquid pipe is connected with the waste liquid container (such as waste liquid bag (6))
- the described liquid replenishment pipe is connected with a plurality of liquid replenishment containers (such as liquid replenishment bag) successively through a plurality of branch pipes, and each A valve is arranged on the branch pipe.
- the number of the rehydration container (eg, rehydration bag) is 1, 2, 3, 4, 5, 6, 7 or 8.
- the number of the rehydration container (such as a rehydration bag) is two, and two different rehydration containers (such as a rehydration bag) contain the rehydration component 1 and the rehydration component 2 respectively.
- the rehydration component 1 includes compound electrolyte injection and human serum albumin aqueous solution
- the rehydration component 2 includes CS10 frozen storage solution.
- the compound electrolyte injection includes 3-7 parts by weight of sodium chloride, 3-7 parts by weight of sodium gluconate, 2-6 parts by weight of sodium acetate, 0.1-0.8 parts by weight of potassium chloride, 0.1-0.6 parts by weight Parts of magnesium chloride and 950-1050 parts by weight of water.
- the concentration of human serum albumin in the aqueous solution of human serum albumin is 15-25% (v/v), preferably 18-22% (v/v).
- the volume ratio of the compound electrolyte injection to the human serum albumin aqueous solution is 80-120:1, preferably 90-110:1, more preferably 95-105:1.
- the CS10 cryopreservation solution includes DMSO aqueous solution.
- the CS10 cryopreservation solution includes 8-12% (v/v) DMSO aqueous solution.
- the cell sample container (such as a cell sample bag) contains cell samples.
- the present invention also provides a method for subpackaging cells with a fully enclosed and fully automated cell subpackaging device as described in the present invention, said method comprising the steps of:
- the cell sample container (such as a cell sample bag) contains a cell sample
- two different rehydration containers (such as a rehydration bag) respectively contain a rehydration component 1 and a rehydration component 2, wherein,
- the rehydration component 1 includes compound electrolyte injection and human serum albumin aqueous solution, and the rehydration component 2 includes CS10 frozen storage solution;
- the cell sample in the cell sample container (such as a cell sample bag) enters the extraction device through the injection tube and the extraction tube, and then the cell sample is discharged through the extraction tube and the extraction device.
- the waste liquid tube enters into the waste liquid container (such as waste liquid bag), so as to perform pipeline rinsing on the sampling tube, extraction tube and waste liquid tube;
- the cell sample in the cell sample container (such as a cell sample bag) enters the extraction device through the injection tube and the extraction tube, and then the cell sample is discharged through the extraction tube and The liquid outlet pipe enters into the cell freezing container (such as a cell freezing bag);
- the rehydration component 1 in the rehydration container (such as a rehydration bag) enters the withdrawal device through the rehydration tube and the extraction tube, and then the rehydration component 1 passes through the extraction tube and the extraction tube through the discharge of the extraction device.
- the liquid outlet pipe enters into the cell freezing container (such as a cell freezing bag);
- the rehydration component 2 in the rehydration container (such as a rehydration bag) enters the withdrawal device through the rehydration tube and the extraction tube, and then through the discharge of the extraction device, the rehydration component 2 passes through the extraction tube and the liquid outlet tube into the cell freezing container (such as a cell freezing bag);
- step (4) the rehydration component 1 and the rehydration component 2 enter into the cell freezing container (such as a cell freezing bag) sequentially, and the volume ratio of the rehydration component 1 and the rehydration component 2 is 1:1.
- the cell freezing container such as a cell freezing bag
- the compound electrolyte injection includes 3-7 parts by weight of sodium chloride, 3-7 parts by weight of sodium gluconate, 2-6 parts by weight of sodium acetate, 0.1-0.8 parts by weight of sodium chloride Potassium, 0.1-0.6 parts by weight of magnesium chloride and 950-1050 parts by weight of water.
- the method further includes step (5): counting the cells in the cell freezing container (such as a cell freezing bag) described in step (4), if the cell density is relatively large, according to the above Replenish the liquid in the step (4). If the cell density is low, supplement the cell sample according to the above step (2), so as to obtain the cell suspension with the required cell density.
- the cell freezing container such as a cell freezing bag
- the method also includes a packing step (6), and the step includes: carrying out a cell freezing container (such as a cell freezing bag) containing the cell suspension with a sampling tube Connect, and the waste liquid pipe is connected with the sub-packaging bag;
- a cell freezing container such as a cell freezing bag
- the cell suspension in the cell freezing container (such as a cell freezing bag) enters the extraction device through the injection tube and the extraction tube, and then the cell suspension passes through the extraction tube and the waste through the discharge of the extraction device.
- the liquid pipe enters into the subpackaging bag, and multiple subpackaging of the cell suspension is realized by changing the subpackaging bag.
- the subpackaging step (6) is after the step (5).
- the fully enclosed and fully automated cell subpackaging device and method of the present invention can perform cell subpackaging under closed conditions, reducing the risk of infection from contact with the external environment.
- the device and method for packaging cells can ensure the survival rate of the subpackaged cells, and the precision of the subpackaged cells is high, thereby improving the quality of the subpackaged cells.
- the cell preparation method of the present invention can quickly prepare immune cells, reduces the cost of the enterprise, improves production capacity, and is suitable for industrial production. Meanwhile, the immune cells prepared by the cell preparation method of the present invention have high quality and can ensure clinical efficacy.
- the amount of positive T cells in the final product bag is 150 ⁇ 10 6 cells, and the frozen storage volume is 40ml; the amount of positive T cells in the cell bag for detection is 37.5 ⁇ 10 6 cells, and the frozen storage volume is 10ml;
- the positive rate was 20.71%; the theoretical cryopreservation density was 19.918 ⁇ 10 6 cells/ml.
- Component 1 Add the HSA (human serum albumin) aqueous solution with a concentration of 20% (v/v) to the compound electrolyte injection (every 1000ml of water contains 5.26g of sodium chloride, 5.02g of sodium gluconate, and 5.02g of sodium acetate) 3.68g, potassium chloride 0.37g, magnesium chloride 0.30g), and then filled into the component 1 bag; wherein the volume ratio of 20% (v/v) HSA aqueous solution and compound electrolyte injection is 1:100.
- HSA human serum albumin
- Component 2 Fill the CS10 cryopreservation solution into the component 2 bag, wherein the CS10 cryopreservation solution is DMSO aqueous solution, and the volume fraction of DMSO is 10%.
- Kit CT-49.1 a fully enclosed and fully automated device for distributing cells
- open Kit CT-49.1 a fully enclosed and fully automated device for distributing cells
- close the clips use a sterile pipette to connect the harvested cell sample bags to position 5, and the 250ml cell cryopreservation bags to At position 7, component 1 bag is at position 8-1, component 2 bag is at position 8-2, and waste liquid bag is at position 6.
- Figure 1 The schematic diagram of the pipeline position of Kit CT-49.1 is shown in Figure 1;
- i. Cell counting After concentrating the cell suspension, connect a 5ml syringe to a 250ml cryopreservation bag containing the cell suspension, shake the cryopreservation bag well, and extract about 0.2ml of the cell suspension for cell counting;
- the volume of component 1 to be added the volume of the cell suspension in the cryopreservation bag + the volume of component 1 to be added;
- Pipeline rinsing input component 1 rinsing volume 10ml, open the pipe clamp, and start the program;
- Pipeline rinsing input component 2 rinsing volume 10ml, open the pipe clamp, and start the program;
- Rinse the pipeline input the rinse volume of the final product as 10ml, open the pipeline clamp, and start the program;
- ii. Cell bag dispensing for detection Before dispensing, first calculate the dispensing volume according to the amount of cells filled in the cell bag for testing and the freezing storage density, then connect the 50ml cell freezing storage bag to position 6, and input the calculated dispensing volume , click " ⁇ " to open the pipeline clamp and start the program. Repeat this step twice to obtain three bags of cell bags for testing;
- Dispensing of the final product Before dispensing, first calculate the dispensing volume according to the amount of cells filled in the final product and the frozen storage density of the final product, then connect the 250ml cell freezing bag to position 6, input the calculated dispensing volume, and click " ⁇ ", open the pipe clamp and start the program. Get a bag of final product;
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Abstract
Description
组分名称 | 分装体积(ml) |
组分1 | 54.7 |
组分2 | 54.7 |
Claims (11)
- 一种全封闭和全自动化的分装细胞的装置,其特征在于,所述的装置包括进样模块、补液模块和液体分配模块;所述的进样模块包括细胞样品容器;所述的补液模块包括多个补液器,所述的补液器包括多个补液容器;所述的液体分配模块包括液体分配器,所述的液体分配器设有进样管、出液管、抽出管、废液管和补液管相连,所述的进样管、出液管、抽出管、废液管和补液管是相通的,且所述的进样管、出液管、抽出管、废液管和补液管上均设有阀门;所述的进样管与所述的细胞样品容器相连,所述的出液管与细胞冷冻容器相连,所述的抽出管与抽出装置相连,所述废液管与废液容器相连,所述的补液管通过多个分支管依次与多个补液容器连接,且各个分支管上设有阀门。
- 如权利要求1所述的装置,其特征在于,所述补液容器的个数为2个,2个不同的补液容器分别含有补液组分1和补液组分2,其中,所述补液组分1包括复方电解质注射液和人血清白蛋白水溶液,所述补液组分2包括CS10冻存液。
- 如权利要求2所述的装置,其特征在于,所述的复方电解质注射液包括3-7重量份氯化钠、3-7重量份葡萄糖酸钠、2-6重量份醋酸钠、0.1-0.8重量份氯化钾、0.1-0.6重量份氯化镁和950-1050重量份水。
- 如权利要求2所述的装置,其特征在于,所述的人血清白蛋白水溶液中的人血清白蛋白的浓度为15-25%(v/v)。
- 如权利要求1所述的装置,其特征在于,所述的抽出装置能够吸入液体和推出液体。
- 如权利要求2如权利要求1所述的装置,其特征在于,所述的复方电解质注射液与所述人血清白蛋白水溶液的体积比为80-120:1,较佳地90-110:1,更佳地95-105:1。
- 如权利要求2所述的装置,其特征在于,所述的CS10冻存液包括8-12%(v/v)DMSO水溶液。
- 一种用如权利要求1所述的装置制备细胞悬液的方法,其特征在于,所述的方法包括步骤:(1)所述的细胞样品容器含有细胞样品,2个不同的补液容器分别含有补 液组分1和补液组分2,其中,所述补液组分1包括复方电解质注射液和人血清白蛋白水溶液,所述补液组分2包括CS10冻存液;(2)通过抽出装置的抽吸使细胞样品容器中的细胞样品经进样管和抽出管进入到抽出装置中,然后再通过抽出装置的排出使细胞样品经抽出管和废液管进入到废液容器中,从而对进样管、抽出管和废液管进行管路润洗;(3)通过抽出装置的抽吸使细胞样品容器中的细胞样品经进样管和抽出管进入到抽出装置中,然后再通过抽出装置的排出使细胞样品经抽出管和出液管进入到细胞冷冻容器中;(4)通过抽出装置的抽吸使得补液容器中补液组分1经补液管和抽出管进入到抽出装置中,然后再通过抽出装置的排出使补液组分1经抽出管和出液管进入到细胞冷冻容器中;通过抽出装置的抽吸使得补液容器中补液组分2经补液管和抽出管进入到抽出装置中,然后再通过抽出装置的排出使补液组分2经抽出管和出液管进入到细胞冷冻容器中;在步骤(4)中,补液组分1和补液组分2依次进入到细胞冷冻容器中,且补液组分1和补液组分2的体积比为1:1。
- 如权利要求8所述的方法,其特征在于,所述的方法还包括步骤(5):对步骤(4)所述的细胞冷冻容器的细胞进行计数,如果细胞密度较大,按照上述步骤(4)的步骤进行补液,如果细胞密度较小,按照上述步骤(2)的步骤进行补充细胞样品,从而得到所需细胞密度的细胞悬液。
- 如权利要求8所述的方法,其特征在于,所述的方法还包括分装步骤(6),所述的步骤包括:将含有所述细胞悬液的细胞冷冻容器与进样管进行连接,所述废液管与分装袋进行连接;通过抽出装置的抽吸使细胞冷冻容器中的细胞悬液经进样管和抽出管进入到抽出装置中,然后再通过抽出装置的排出使细胞悬液经抽出管和废液管进入到分装袋中,通过更换分装袋实现细胞悬液的多次分装。
- 如权利要求8所述的方法,其特征在于,所述的复方电解质注射液包括3-7重量份氯化钠、3-7重量份葡萄糖酸钠、2-6重量份醋酸钠、0.1-0.8重量份氯化钾、0.1-0.6重量份氯化镁和950-1050重量份水。
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