Protein-free cell cryopreservation liquid and application thereof

A cryopreservation solution, protein-free technology, applied in the field of cell cryopreservation, can solve the problems of cumbersome cryopreservation operation, unclear composition of cryopreservation solution, poor safety, etc., and achieves high resuscitation rate, simple and clear components, and easy to use. Effect

Inactive Publication Date: 2020-10-30
苏州依科赛生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The present invention provides a novel protein-free cell cryopreservation solution and its application, aiming to solve the technical defects of the prior art that the components of the cryopreservation solution are not clear, the safety is poor, and the cryopreservation operation is cumbersome.

Method used

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  • Protein-free cell cryopreservation liquid and application thereof
  • Protein-free cell cryopreservation liquid and application thereof

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Experimental program
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Effect test

Embodiment 1

[0029] Embodiment 1 A kind of protein-free cell cryopreservation liquid

[0030] 1. A protein-free cell freezing solution,

[0031] Among them, DMEM / F12 accounts for 93% of the total volume of the total protein-free cell freezing solution, the concentration of hydroxyethyl starch in the cell freezing solution is 3g / L, and the concentration of threonine in the cell freezing solution is 0.05g / L L, the concentration of polyvinyl alcohol (PVA) in the cell freezing solution is 1g / L, the proportion of DMSO in the cell freezing solution is 5%, and the proportion of raffinose in the cell freezing solution is 1g / L, Propylene glycol accounts for 2% in the cell freezing solution;

[0032] 2. The preparation method of this protein-free cell freezing solution comprises the following steps:

[0033] S1 In a normal working biological safety cabinet, vortex and mix the above-mentioned DMEM / F12 and DMSO in proportion, so that DMEM / F12 accounts for 93% of the total volume of the total protein...

Embodiment 2

[0040] Embodiment 2 A kind of protein-free cell cryopreservation solution

[0041] 1. A protein-free cell cryopreservation solution, wherein DMEM / F12 accounts for 91.5% of the total volume of the total protein-free cell cryopreservation solution, DMSO accounts for 7.5% of the total volume of the total protein-free cell cryopreservation solution, and hydroxyethyl starch in the cell The concentration in the freezing solution is 5g / L, the concentration of threonine in the cell freezing solution is 0.05g / L, the concentration of polyvinyl alcohol (PVA) in the cell freezing solution is 1.5g / L, and DMSO is in The proportion of the cell freezing solution is 7.5%, the proportion of raffinose in the cell freezing solution is 1g / L, and the proportion of propylene glycol in the cell freezing solution is 1%;

[0042] 2. The preparation method of this protein-free cell freezing solution comprises the following steps:

[0043] S1 In a normal working biological safety cabinet, vortex and mix t...

Embodiment 3

[0050] Embodiment 3 A kind of protein-free cell cryopreservation solution

[0051] 1. A protein-free cell cryopreservation solution, wherein DMEM / F12 accounts for 90% of the total volume of the total protein-free cell cryopreservation solution, DMSO accounts for 10% of the total volume of the total protein-free cell cryopreservation solution, and hydroxyethyl starch in the cell The concentration in the freezing solution is 3 / L, the concentration of threonine in the cell freezing solution is 0.05g / L, the concentration of polyvinyl alcohol (PVA) in the cell freezing solution is 2g / L, and the concentration of DMSO in the cell freezing solution is 2g / L. The proportion in the cryopreservation solution is 10%, the proportion of raffinose in the cell cryopreservation solution is 0.5g / L, and the proportion of propylene glycol in the cell cryopreservation solution is 0.1%;

[0052] 2. The preparation method of this protein-free cell freezing solution comprises the following steps:

[...

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Abstract

The invention relates to a novel cell cryopreservation liquid and application thereof, and discloses a protein-free cell cryopreservation liquid which comprises DMEM / F12, threonine, hydroxyethyl starch, raffinose, polyvinyl alcohol (PVA), dimethyl sulfoxide (DMSO) and propylene glycol. The protein-free cell cryopreservation liquid is free of protein components and animal-derived components, clearin chemical components and universally suitable for ultralow-temperature storage of various cell types or tissues. The invention further provides a using method of the protein-free cell cryopreservation liquid, and the protein-free cell cryopreservation liquid can be used for programmed freezing and is also suitable for a non-programmed freezing mode. The protein-free cell cryopreservation liquidhas clear and stable components, eliminates instability of serum-containing cryopreservation liquid, solves the problem that traditional serum-containing cryopreservation liquid possibly introduces pathogenic bacteria, does not contain protein components, eliminates body immune response possibly caused by foreign protein, has higher safety and is particularly suitable for the field of clinical application. The cells preserved by the cryopreservation liquid can be recovered after being cryopreserved for one month in a short term, the cell viability can be kept at 95% or above, the cell viability can still reach 90% or above after the cells are recovered after being preserved for one year, and the cell viability and functions are not affected.

Description

technical field [0001] The invention belongs to the technical field of cell culture, in particular to a cell cryopreservation solution. Background technique [0002] Cell cryopreservation is to store cells in a low temperature environment, so that cells can temporarily escape from the growth state and maintain cell characteristics. The cost of storing cells by freezing is low, and they can be revived and continued to be cultured when needed. At the same time, cryopreservation can also avoid the loss of cell species caused by cell contamination during culture. [0003] Cell cryopreservation solution is a protective reagent that must be used when cells are cryopreserved. Cells are protected to prevent or reduce damage to cells caused by ice crystals during freezing. [0004] The traditional cell cryopreservation solution is prepared by adding fetal bovine serum and osmotic protectant DMSO to the basal medium, or using 90% fetal bovine serum and adding 10% DMSO, but this kin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 于宝利孙芳陈旭陈刚李其雷杨建国
Owner 苏州依科赛生物科技股份有限公司
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