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NK cell culture kit and usage method thereof and NK cell obtained therefrom

A technology for NK cells and cell culture, applied in the field of NK cells, can solve the problem of no induction kit, achieve the effect of simple culture and enhanced killing activity

Inactive Publication Date: 2017-08-18
TIANJIN PURUI SAIER BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no simple, efficient and stable induction kit

Method used

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  • NK cell culture kit and usage method thereof and NK cell obtained therefrom
  • NK cell culture kit and usage method thereof and NK cell obtained therefrom
  • NK cell culture kit and usage method thereof and NK cell obtained therefrom

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] (1) Take the T-75 cell culture bottle one day before the experiment, add 5ml serum-free lymphocyte medium (Lonza company serum-free lymphocyte medium, article number: 04-418Q), a pretreatment factor NK-A, including Cultured flask, overnight at 4°C;

[0092] (2) Take two mononuclear lymphocyte separation centrifuge tubes, add 15ml of lymphocyte separation solution along the reserved sample port into each mononuclear lymphocyte separation centrifuge tube, Add 30ml of routine peripheral blood collected, Ficoll-Hypaque density gradient centrifugation, 2000rpm / min, centrifugation for 23min;

[0093] (3) Collect the upper layer of plasma in step (2), put the collected plasma in a water bath at 56°C for 30 minutes to inactivate complement; centrifuge at 3000 rpm / min for 8 minutes to remove the precipitates in the plasma, and transfer the upper layer of autologous serum to a centrifuge tube, A total of 26ml, placed in a 4 ℃ refrigerator for later use;

[0094] (4) Collect the...

Embodiment 2

[0104] (1) Take the T-75 cell culture bottle one day before the experiment, add 5ml serum-free lymphocyte medium (Lonza company serum-free lymphocyte medium, article number: 04-418Q), a pretreatment factor NK-A, including Cultured flask, overnight at 4°C;

[0105] (2) Take two mononuclear lymphocyte separation centrifuge tubes, add 15ml of lymphocyte separation solution along the reserved sample port into each mononuclear lymphocyte separation centrifuge tube, Add 30ml of routine peripheral blood collected, Ficoll-Hypaque density gradient centrifugation, 2000rpm / min, centrifugation for 23min;

[0106](3) Collect the upper layer of plasma in step (2), put the collected plasma in a water bath at 56°C for 30 minutes to inactivate complement; centrifuge at 3000 rpm / min for 8 minutes to remove the precipitates in the plasma, and transfer the upper layer of autologous serum to a centrifuge tube, A total of 24ml, placed in a 4 ℃ refrigerator for later use;

[0107] (4) Collect the ...

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Abstract

The invention discloses an NK cell culture kit and a usage method thereof and an NK cell obtained therefrom. The kit comprises an NK kit set and an NK consumable kit set, wherein the consumable kit set consists of a mononuclear lymphocyte cell separation centrifugal tube, a portable cell sample adding pre-filling stand, a cell culture bag tray and a cell culture bag; and the reagent box set consists of a pretreatment factor NK-A, a pretreatment factor NK-B, an amplification factor NK-C, an amplification factor NK-D, an amplification factor NK-E and a serum-free lymphocytes cell culture medium. More than 2*10<9> lymphocyte cells can be amplified within short time by matching the NK culture kit with basic experimental consumables, wherein the ratio of the NK cells (CD3-CD56+) can achieve more than 60%, thus providing convenience for amplifying NK cells in vitro on a large scale and conducting related studies and clinical trials.

Description

technical field [0001] The invention relates to the field of cell biotechnology, in particular to a set of NK cell culture kits capable of standardized production operations, a use method thereof and obtained NK cells. Using this kit and basic experimental consumables can conveniently expand a large number of NK cells in a short period of time, providing convenience for large-scale expansion of NK cells in vitro and related research and clinical trials. Background technique [0002] In 2000, the International Tumor Biotherapy and Gene Therapy Annual Conference pointed out that "immune cell therapy is the only possible way to completely eliminate cancer cells in modern technology". Dreams are turning into reality, and cancer is being gradually defeated by human beings. Cellular immunotherapy is the fourth largest cancer treatment mode after surgery, radiotherapy, and chemotherapy. As a hotspot in the clinical research of immunotherapy, NK cells have different induction and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0646C12N2501/2302C12N2501/2312C12N2501/2315C12N2501/599
Inventor 熊亮鲁振宇张冰晶韩洪起秦臻高婧彧徐悦黄文敬
Owner TIANJIN PURUI SAIER BIOLOGICAL TECH CO LTD
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