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Isolated culture method of high-purity NK cells

A technology of NK cells and culture methods, applied in the field of separation and culture of high-purity NK cells, can solve problems such as poor quantity and purity, and achieve the effects of avoiding amplification and killing activities, reducing culture costs, and increasing the number of cells

Pending Publication Date: 2021-07-02
SICHUAN NEO LIFE STEM CELL BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In the in vitro culture method of NK cells reported in the patent application with publication number CN 103627672A, in the obtained culture products, NK cells are only amplified by 102.81±94.06 times, and the traditional Ficoll density gradient centrifugation method is used to separate NK cells, and the quantity and purity are not high. good

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  • Isolated culture method of high-purity NK cells
  • Isolated culture method of high-purity NK cells
  • Isolated culture method of high-purity NK cells

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Embodiment 1

[0052] Embodiment 1 The separation culture method of the present invention

[0053] 1. Isolation and culture of NK cells

[0054] 1. Isolation of peripheral blood / cord blood mononuclear cells and NK cell sorting

[0055] Peripheral blood collection: 30-60 mL of autologous peripheral blood from tumor patients or healthy volunteers;

[0056] Umbilical cord blood collection: 70-90 mL of cord blood from healthy full-term fetuses;

[0057] Use sodium citrate, or sodium heparin anticoagulant blood collection tubes or blood collection bags to collect fresh blood, and the time from collection to NK separation and preparation should not exceed 24 hours.

[0058] Wipe the whole blood collection tube with 75% alcohol, pour the anticoagulated whole blood into a 50ml centrifuge tube and mix well, and take 0.5mL anticoagulated blood for cell five-differential counting and flow cytometry detection. Centrifugation conditions: centrifugal force 800g, centrifuge for 8min, absorb the supernat...

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Abstract

The invention provides a kit for isolated culture of high-purity NK cells, the kit comprises a culture medium 1-3, and the invention also provides an isolated culture method of the high-purity NK cells. By utilizing the kit and the isolated culture method provided by the invention, the NK cells meeting clinical use can be efficiently obtained, the number of the cells is large, the purity is high, the killing activity is good, and the kit and the isolated culture method have outstanding advantages in the aspect of allogeneic NK feedback and have a good application prospect.

Description

[0001] The present invention is an application date: September 5, 2017, application number: 201710791174.8, patent name: a division of a method for separating and culturing high-purity NK cells. technical field [0002] The invention belongs to the field of cell biology, and in particular relates to a method for separating and culturing high-purity NK cells. Background technique [0003] Natural killer cells (NK) are important immune cells of the body. As the first line of defense of the body's defense system, they can not only exert their main tumor-killing effect in the natural immune system, but also secrete different Cytokines and various chemokines regulate the body's acquired immune response and are indispensable effector cells for the body to exert immune effects. Compared with αβT cells, γδT cells, CIK and other cells used for immunotherapy, NK cells have received more attention and attention due to their high cell killing activity, rapid onset of action, and not bei...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0646C12N2509/00C12N2501/2302C12N2501/2315
Inventor 杨本艳姿李雪莲陈强
Owner SICHUAN NEO LIFE STEM CELL BIOTECH