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Serum-free cell cryopreservation solution as well as preparation method and application thereof

A cryopreservation, serum-free technology, applied in the field of cell culture, can solve the problems of cell body damage, increase pollution, and time-consuming, and achieve the effects of reducing cell aggregation, improving survival rate, and simple operation.

Active Publication Date: 2019-08-02
苏州博特龙免疫技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But there are still some defects: DMSO has certain toxicity, and high concentration of DMSO has great toxicity, which will cause damage to the cell body; FBS is an animal-derived substance, which may carry the risk of animal-derived viruses, and human serum may also contain Human-originated viruses have potential risks in clinical application and also increase the chance of contamination
However, there are still some defects, such as: the survival rate and activity of cells after recovery still need to be improved; the toxicity of serum-free cell cryopreservation agent is high; The rate is different, and the freezing rate has a greater impact on the freezing of cells, the freezing process is complicated, takes a long time, and the freezing effect is unstable, etc.

Method used

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  • Serum-free cell cryopreservation solution as well as preparation method and application thereof
  • Serum-free cell cryopreservation solution as well as preparation method and application thereof
  • Serum-free cell cryopreservation solution as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: A kind of serum-free cell cryopreservation medium

[0031] 1. A serum-free cell freezing solution comprises: serum-free DMEM low-sugar medium 83% (v / v), dimethylsulfoxide 10% (v / v), hydroxyethyl starch 2% (w / v) , Catechin 2% (w / v), Sodium Tetraborate 0.2% (w / v), Sodium Hyaluronate 2% (w / v) and Vitamin C 0.8% (w / v).

[0032] The mass concentration ratio of hydroxyethyl starch and catechin is 1:1; the mass concentration ratio of sodium tetraborate and sodium hyaluronate is 1:10.

[0033] 2. The preparation method of this serum-free cell cryopreservation solution comprises the following steps:

[0034] (1) In a sterile environment, mix serum-free DMEM low-sugar medium and dimethyl sulfoxide according to the ratio, and stir at room temperature for 5 minutes, so that serum-free DMEM low-sugar medium and dimethyl sulfoxide can be frozen in serum-free cells The final concentrations in the solution were 83% (w / v) and 10% (v / v) respectively, and they were mixed u...

Embodiment 2

[0039] Embodiment 2: A kind of serum-free cell cryopreservation medium

[0040] 1. A serum-free cell freezing solution comprises: serum-free DMEM high-glucose medium 84% (v / v), dimethyl sulfoxide 10% (v / v), hydroxyethyl starch 3% (w / v ), catechin 1% (w / v), sodium tetraborate 0.4% (w / v), sodium hyaluronate 1% (w / v) and vitamin C 0.6% (w / v).

[0041] The mass concentration ratio of hydroxyethyl starch and catechin is 3:1; the mass concentration ratio of sodium tetraborate and sodium hyaluronate is 4:10.

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

[0043] (1) In a sterile environment, mix the serum-free DMEM high-glucose medium and dimethyl sulfoxide according to the ratio, and stir at room temperature for 5 minutes to make the serum-free DMEM high-glucose medium and dimethyl sulfoxide in serum-free cells The final concentrations in the frozen storage solution were 84% (w / v) and 10% (v / v), and mixed evenly...

Embodiment 3

[0048] Embodiment 3: A kind of serum-free cell cryopreservation medium

[0049] 1. A serum-free cell freezing solution comprises: serum-free 1640 medium 85% (v / v), dimethyl sulfoxide 10% (v / v), hydroxyethyl starch 2.5% (w / v), Catechin 0.5% (w / v), Sodium Tetraborate 0.5% (w / v), Sodium Hyaluronate 1% (w / v) and Vitamin C 0.5% (w / v).

[0050] The mass concentration ratio of hydroxyethyl starch and catechin is 5:1; the mass concentration ratio of sodium tetraborate and sodium hyaluronate is 5:10.

[0051] 2. The preparation method of this serum-free cell cryopreservation solution comprises the following steps:

[0052] (1) In a sterile environment, mix the serum-free 1640 medium and dimethyl sulfoxide in proportion, and stir at room temperature for 5 minutes to make the serum-free 1640 medium and dimethyl sulfoxide in the serum-free cell freezing solution The final concentrations were 85% (w / v) and 10% (v / v) respectively, mixed uniformly to prepare mixed solution 1;

[0053] (2)...

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PUM

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Abstract

The invention provides a serum-free cell cryopreservation solution as well as a preparation method and application thereof, and relates to the technical field of cell culture. The cryopreservation solution comprises a serum-free medium, dimethyl sulfoxide, hydroxyethyl starch, catechin, sodium tetraborate, sodium hyaluronate and vitamin C, and does not contain bovine serum or other protein components, thereby avoiding the introduction of exogenous factors causing cell seed contamination. The cryopreservation solution has low toxicity and remarkable cryopreservation effect, and the cell survival rate after freezing for 3 months reaches 95% or above. In addition, a use method of the serum-free cell cryopreservation solution is simple to operate, no complicated program for cooling is required, and the serum-free cell cryopreservation solution can be widely used for cryopreservation of various mammalian cells.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a serum-free cell cryopreservation solution and its preparation method and application. Background technique [0002] Cell cryopreservation is one of the main methods of cell preservation, and its purpose is to slow down the metabolic activity of cells and maintain life in a low temperature environment. During the freezing of cell suspensions, the formation of ice crystals is one of the main causes of cell death. The main factors affecting cell cryopreservation include cell cryopreservation medium and cryopreservation rate. As a solution that must be used for cell cryopreservation, cell cryopreservation solution is used to suspend cells that need to be cryopreserved, supply nutrients needed for cell life metabolism, and prevent or reduce damage to cells caused by frozen ice crystals. There are two effects of freezing rate on cell survival rate, one is damage under slow fre...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/021A01N1/0226
Inventor 刘立成
Owner 苏州博特龙免疫技术有限公司
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