Cryopreservation solution for cultured NKT cells and preparation method of cryopreservation solution

A technology of NKT cells and cryopreservation solution, applied in the field of cell cryopreservation solution, can solve problems such as adverse reactions, pollution and allergens, and affect treatment results, and achieve high safety, maintain activity, and good cryopreservation 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

[0006] In the prior art, cryopreservation solutions prepared by adding different proportions of fetal bovine serum (FBS) and dimethyl sulfoxide (DMSO) are usually used, and then programmed cooling at different temperatures and times is used to freeze the cultured NKT cells. Adding DMSO is just to prevent the formation of intracellular ice crystals from damaging cells during freezing, and adding FBS is to protect cells from DMSO damage and provide nutrition, but FBS is a heterogeneous substance with complex components and introduces pollution and allergen risk, not suitable for clinical application
Especially in cell therapy, the presence of heterologous proteins may cause unknown adverse reactions and seriously affect the treatment results

Method used

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  • Cryopreservation solution for cultured NKT cells and preparation method of cryopreservation solution
  • Cryopreservation solution for cultured NKT cells and preparation method of cryopreservation solution
  • Cryopreservation solution for cultured NKT cells and preparation method of cryopreservation solution

Examples

Experimental program
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Embodiment 1

[0030] This embodiment provides a method for preparing a frozen storage solution of cultured NKT cells, comprising the following steps:

[0031] 1) Add lentinan with a concentration of 1-6 mg / mL and seaweed polysaccharide with a concentration of 0.05-0.2 mg / mL into autologous serum respectively, then slowly add DMSO, and add lentinan: seaweed polysaccharide: autologous serum: DMSO according to the volume ratio 1~3:1~3:1~5:1, put the prepared solution A in the refrigerator at 2~8℃ to pre-cool, and set aside;

[0032] 2) Then take out the above solution A, slowly add it to the autologous serum, add solution A: autologous serum ratio of 1-3:1 by volume percentage, and prepare NKT cell cryopreservation solution.

Embodiment 2

[0034] In this example, the cryopreservation solution of cultured NKT cells was prepared according to the method described in Example 1. The cryopreservation solution was prepared from solution A and autologous serum, and the volume ratio of solution A and autologous serum was 1:1. in:

[0035] The solution A includes lentinan, seaweed polysaccharide, autologous serum and DMSO, wherein the volume ratio of lentinan, seaweed polysaccharide, autologous serum and DMSO is 1:1:1:1, wherein the concentration of lentinan is 1 mg / mL, and the seaweed The polysaccharide concentration was 0.05 mg / mL.

Embodiment 3

[0037] In this example, a cryopreservation solution of cultured NKT cells was prepared according to the method described in Example 1. The cryopreservation solution was prepared from solution A and autologous serum, and the volume ratio of solution A and autologous serum was 2:1. in:

[0038] The solution A includes lentinan, seaweed polysaccharide, autologous serum and DMSO, wherein the volume ratio of lentinan, seaweed polysaccharide, autologous serum and DMSO is 2:2:3:1, wherein the concentration of lentinan is 6 mg / mL, and the seaweed The polysaccharide concentration was 0.2 mg / mL.

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Abstract

The invention relates to the technical field of cell cryopreservation solutions, and in particular relates to a cryopreservation solution for the cultured NKT cells and a preparation method of the cryopreservation solution. The cryopreservation solution for the cultured NKT cells comprises lentinan, algal polysaccharides, autoserum and DMSO. By virtue of adding the autoserum, the pollution of foreign animal viruses and the introduction of foreign proteins are avoided, and the safety is high; by virtue of adding the lentinan and the algal polysaccharides, the activity of the cells can be effectively 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 cryopreservation liquid, in particular to a cryopreservation liquid of cultured NKT cells 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. During cryopreservation at ultra-low temperature, 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 stopped, thus maintaining cell activity. [0003] In recent years, research on NKT cell phe...

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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