Construction method of CIK cell bank

A construction method and cell technology, applied in the field of cell bank construction, can solve problems such as difficulty in meeting the needs of use

Pending Publication Date: 2021-09-28
海南优尼科尔生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as the age increases, the activity of CIK cells in the human body will gradually decrease, and it will be difficult to meet the needs of use when needed. It becomes a new choice to store the cells while they are young.

Method used

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  • Construction method of CIK cell bank

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Human Peripheral CIK cell bank construction method, comprising the steps of:

[0069] 1) collected from healthy donor peripheral blood under the 100ml bag containing sterile conditions to a sodium citrate anticoagulant was transferred to a 50ml centrifuge tube, centrifuged by using a centrifuge, centrifugation conditions: 4 ℃, 2500 rpm, 10min , 8 l 8 down, the upper layer as a pale yellow liquid to a new 50ml centrifuge tube 56 is placed deg.] C water bath inactivated 30min, centrifuged using a centrifuge, centrifugation conditions: 4 ℃, 3000 rpm, centrifugal 8min, 8 liters 8 down, the supernatant was collected, namely autologous plasma, placed in -20 ℃ refrigerator spare;

[0070] 2) Peripheral blood mononuclear cells: the lower layer is removed after the red liquid volume of autologous plasma according to the meter-fold diluted with physiological saline and mix by pipetting, added 6wt% of which 0.5 volumes of a solution of hydroxyethyl starch, after the mixing was allowed...

Embodiment 2

[0076] Human Peripheral CIK cell bank construction method, comprising the steps of:

[0077] 1) collected from healthy donor peripheral blood under the 100ml bag containing sterile conditions to a sodium citrate anticoagulant was transferred to a 50ml centrifuge tube, centrifuged by using a centrifuge, centrifugation conditions: 4 ℃, 2500 rpm, 10min , 1 drop 1 liter, the upper layer as a pale yellow liquid to a new 50ml centrifuge tube 56 is placed deg.] C water bath inactivated 30min, centrifuged using a centrifuge, centrifugation conditions: 4 ℃, 3000 rpm, centrifuge 5min, 9 liters 9 down, the supernatant was collected, namely autologous plasma, placed in -20 ℃ refrigerator spare;

[0078] 2) Peripheral blood mononuclear cells: the lower layer is removed after the red liquid volume of autologous plasma according to the meter-fold diluted with physiological saline and mix by pipetting, the mixture was added slowly to a 20ml centrifuge containing 20ml of lymphocyte separation medi...

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Abstract

The invention discloses a construction method of a CIK cell bank. The construction method sequentially comprises the following steps: collecting human peripheral blood, separating autologous plasma, separating peripheral blood mononuclear cells, carrying out multiplication culture on CIK cells, detecting the CIK cells, cryopreserving the CIK cells, coding, warehousing and uploading cell information files. The invention belongs to the technical field of cell bank construction, and particularly relates to the construction method based on the CIK cell bank.

Description

Technical field [0001] The present invention belongs to the field of cell library construction, and is specifically a method of constructing a CIK cell bank. Background technique [0002] Cells in the 1980s have brought new hopes to patients with tumor patients. Cytokine-Induced Killer (CIK) is a group of CD3 + and CD56 + T cells in combination with various cytokines (IFN-γ, IL-2, IL-1, etc.), anti-CD3 antibody Heterogeneous cells exhibit anti-tumor effects through cytotoxiclymphocyte, CTL) lysolar effect and cell apoptosis, and eliminate micro residual lesions, prevent cancer cell proliferation and tumor recurrence, is a new type Main functional cells in overhaul immunotherapy. In physiological state, expressing CD3 + and CD56 + T lymphocytes accounted for (1-5)% in normal human peripheral blood, far from being able to meet the needs of clinical treatment. With the increasingly perfect in vitro amplification technology, it is a reality for clinical supply of sufficient CD3 + and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C40B50/06C12N5/0783A01N1/02
CPCC40B50/06C12N5/0638A01N1/0221C12N2509/00C12N2501/2302C12N2501/24C12N2501/515
Inventor 王晓宇李崴郭康合林冠妃李秋宛
Owner 海南优尼科尔生物科技有限公司
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