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Cell freezing medium and application thereof

A technology for cryopreservation and cells, applied in the field of cells, which can solve the problems of bacterial and viral contamination, uncertain composition, etc.

Inactive Publication Date: 2018-05-08
湖南丰晖生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The commonly used cell cryopreservation scheme is to use DMSO as the cryoprotectant and add bovine serum at the same time. This scheme can be used for the preservation of scientific research products, but there are problems of cytotoxicity, uncertain composition, and possible introduction of pollution sources such as bacteria and viruses.

Method used

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  • Cell freezing medium and application thereof
  • Cell freezing medium and application thereof
  • Cell freezing medium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] 1. Cell cryopreservation solution includes the following components:

[0037]

[0038] 2. Take the cells in the logarithmic growth phase, remove the old culture medium, and wash with PBS.

[0039] 3. Remove the PBS, add an appropriate amount of trypsin (covering the surface of the culture dish) to digest the cells growing in a single layer;

[0040] 4. Centrifuge at 1000rpm for 5min;

[0041] 5. Remove trypsin, add an appropriate amount of prepared frozen culture medium, blow gently with a pipette to make the cells uniform, count, and adjust the final density of cells in the frozen medium to 5×10 6 / ml~1×10 7 / ml;

[0042] 6. Divide the cells into cryopreservation tubes, 1-1.5ml per tube;

[0043] 7. Mark the name of the cells, the time of freezing and the operator on the cryopreservation tube;

[0044] 8. Cryopreservation: The standard freezing procedure is -1°C / min; when the temperature is below -25°C, it can be increased to -10°C / min; when it reaches -100°C, ...

Embodiment 2

[0052] 1. Cell cryopreservation solution includes the following components:

[0053]

[0054] 2. Take the cells in the logarithmic growth phase, remove the old culture medium, and wash with PBS.

[0055] 3. Remove the PBS, add an appropriate amount of trypsin (covering the surface of the culture dish) to digest the cells growing in a single layer;

[0056] 4. Centrifuge at 1000rpm for 5min;

[0057] 5. Remove trypsin, add an appropriate amount of prepared frozen culture medium, blow gently with a pipette to make the cells uniform, count, and adjust the final density of cells in the frozen medium to 5×10 6 / ml~1×10 7 / ml;

[0058] 6. Divide the cells into cryopreservation tubes, 1-1.5ml per tube;

[0059] 7. Mark the name of the cells, the time of freezing and the operator on the cryopreservation tube;

[0060] 8. Cryopreservation: The standard freezing procedure is -2°C / min; when the temperature is below -25°C, it can be increased to -5°C / min; when it reaches -100°C, i...

Embodiment 3

[0068] 1. Cell cryopreservation solution includes the following components:

[0069]

[0070]

[0071] 2. Take the cells in the logarithmic growth phase, remove the old culture medium, and wash with PBS.

[0072] 3. Remove the PBS, add an appropriate amount of trypsin (covering the surface of the culture dish) to digest the cells growing in a single layer;

[0073] 4. Centrifuge at 1000rpm for 5min;

[0074] 5. Remove trypsin, add an appropriate amount of prepared frozen culture medium, blow gently with a pipette to make the cells uniform, count, and adjust the final density of cells in the frozen medium to 5×10 6 / ml~1×10 7 / ml;

[0075] 6. Divide the cells into cryopreservation tubes, 1-1.5ml per tube;

[0076] 7. Mark the name of the cells, the time of freezing and the operator on the cryopreservation tube;

[0077] 8. Cryopreservation: The standard freezing procedure is a cooling rate of -2°C / min; when the temperature is below -25°C, it can be increased to -8°C / ...

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Abstract

The invention relates to the field of cells, in particular to a cell freezing medium and application thereof. The cell freezing medium is prepared by dissolving glycerol, propanediol, trehalose, sucrose, human albumin, glucose and the like in PBS buffer solution, wherein permeable protection fluid comprises the glycerol and the propanediol, and impermeable protection fluid comprises the trehalose,the sucrose and the human albumin, meanwhile the glucose is used for proving energy to cells, and the PBS buffer solution is used for maintaining pH stability of the cell freezing medium. According to the cell freezing medium, components are fixed, batch-to-batch variation is small, and pollutant sources such as germs and viruses are not introduced, the permeable and impermeable liquids are usedsimultaneously to provide frost resisting capability for the cells to improve survival rate of the cells, the glucose is further added to provide energy for the cells to further improve the survival rate of the cells, and finally the PBS buffer solution is used for maintaining the pH stability of the cell freezing medium, so that environment influence on the cells is reduced to improve the survival rate of the cells.

Description

technical field [0001] The invention relates to the field of cells, in particular to a cell cryopreservation solution and its application. Background technique [0002] Cell cryopreservation is a technique in which cells are placed in a low temperature environment to reduce cell metabolism for long-term storage. Cell cryopreservation is one of the main methods of cell preservation, which plays a role in cell preservation. [0003] Cell cryopreservation is one of the main methods of cell preservation. Using cryopreservation technology to store cells in -196°C liquid nitrogen at low temperature can temporarily remove the cells from the growth state and preserve their cell characteristics, so that the cells can be revived for experiments when needed. Moreover, moderately preserving a certain amount of cells can prevent the cells from being lost due to contamination of the cells being cultured or other accidents, which plays a role in cell preservation. In addition, some cell...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 许澎
Owner 湖南丰晖生物科技有限公司
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