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Cryopreservation liquid for cultured NK cells and preparation method of cryopreservation liquid

A technology of NK cells and cryopreservation solution, which is applied in the field of NK cell cryopreservation solution and its preparation, can solve the problems of low NK cell activity and increased risk of pathogen contamination, and achieve good cryopreservation effect, high safety and maintain activity Effect

Active Publication Date: 2016-01-06
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the former method preserves cells for a long time, the cryopreservation solution prepared with fetal bovine serum introduces foreign proteins, which increases the risk of pathogen contamination
The use of autologous serum instead of fetal bovine serum can completely eliminate the risk of infection by exogenous pollutants, but the activity of NK cells recovered and cultured after cryopreservation is low

Method used

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  • Cryopreservation liquid for cultured NK cells and preparation method of cryopreservation liquid
  • Cryopreservation liquid for cultured NK cells and preparation method of cryopreservation liquid
  • Cryopreservation liquid for cultured NK cells and preparation method of cryopreservation liquid

Examples

Experimental program
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Effect test

Embodiment 1

[0032] This embodiment provides a method for preparing a frozen storage solution of cultured NK cells, comprising the following steps:

[0033] 1) Preparation of autologous serum mixture: Weigh appropriate amount of tea polyphenols, grape seed extract and vitamin C and add them to autologous serum to prepare autologous serum mixture;

[0034] 2) Preparation of solution A: Slowly add DMSO to the autologous serum mixture prepared in step 1). The volume ratio of autologous serum mixture to DMSO is 3-6:1 to prepare solution A and place it in 2-8 ℃ refrigerator pre-cooling, spare;

[0035] 3) Take out the pre-cooled solution A, and slowly add it to autologous serum, the volume ratio of solution A to autologous serum is 1:0.5-2, and prepare a cryopreservation solution, in which tea polyphenols, grape seed extract, The final concentrations of vitamin C were 0.5-5 mg / ml, 0.125-0.5 mg / ml, and 0.5-5 mg / ml, respectively.

Embodiment 2

[0037] In this example, the cryopreservation solution of cultured NK cells was prepared according to the method described in Example 1, which was prepared from solution A and autologous serum. The volume ratio of solution A and autologous serum was 1:0.5, wherein:

[0038] Solution A is prepared from autologous serum mixture and DMSO, the volume ratio of autologous serum mixture to DMSO is 3:1;

[0039] The autologous serum mixture includes tea polyphenols, grape seed extract, vitamin C, and autologous serum. The final concentrations of tea polyphenols, grape seed extract, and vitamin C in the autologous serum mixture are 0.5 mg / ml and 0.125 mg / ml respectively. ml, 0.5mg / ml.

Embodiment 3

[0041] In this example, the cryopreservation solution of cultured NK cells was prepared according to the method described in Example 1, which was prepared from solution A and autologous serum. The volume ratio of solution A and autologous serum was 1:2, wherein:

[0042] Solution A is prepared from autologous serum mixture and DMSO, the volume ratio of autologous serum mixture to DMSO is 6:1;

[0043] The autologous serum mixture includes tea polyphenols, grape seed extract, vitamin C and autologous serum, and the final concentrations of tea polyphenols, grape seed extract, and vitamin C in the autologous serum mixture are 5mg / ml and 0.5mg / ml respectively , 5mg / ml.

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Abstract

The invention relates to the technical field of cell cryopreservation liquids, and in particular relates to a cryopreservation liquid for cultured NK cells and a preparation method of the cryopreservation liquid. The cryopreservation liquid for the cultured NK cells comprises tea polyphenol, a grape seed extract, vitamin C and autoserum. By virtue of adopting the autoserum, pollution of exogenous animal viruses and introduction of exogenous proteins are avoided, and the safety is high; by virtue of adding the tea polyphenol, the grape seed extract (procyanidine) and the vitamin C, cell oxidization can be effectively suppressed, the activity of the cells can be retained, and no harm to a human body is generated; meanwhile, the state of the cells recovered from cryopreservation is basically close to the state before cryopreservation, and the cryopreservation effect is good.

Description

technical field [0001] The invention relates to the technical field of cell freezing solution, in particular to a cultured NK cell freezing solution and a preparation method thereof. Background technique [0002] Cell cryopreservation is an effective method for long-term preservation and maintenance of cell activity. Its cryopreservation goes through two stages of low-temperature cryopreservation and ultra-low temperature cryopreservation. Low-temperature cryopreservation can reduce cell metabolism, and ultra-low temperature (in liquid nitrogen at -198°C) Cryopreservation can preserve the metabolism of living cells and almost completely stop cell growth. In ultra-low temperature cryopreservation, the activities of various enzymes that regulate and control cell growth and metabolism are inhibited, so that the biochemical reactions in cells are very slow, or even stop, so as to maintain cell activity. [0003] However, if it is simply frozen, the cells will be damaged in two ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 陈海佳王一飞葛啸虎李丽娟
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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