Cell cryopreservation composition and application thereof

A composition and cell technology, applied in the field of cells, can solve the problems of poor cell survival rate and achieve the effect of improving survival rate, shortening cycle and reducing cost

Active Publication Date: 2018-06-01
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] Most of the existing technologies use DMSO or glycerin as protective agents for cryopreservation, but the survival rate of cells after recovery is not very good, usually around 50%.

Method used

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  • Cell cryopreservation composition and application thereof
  • Cell cryopreservation composition and application thereof
  • Cell cryopreservation composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] (1) Cell cryopreservation:

[0035] Freezing solution formula:

[0036]

[0037] 1. Prepare cell freezing culture medium, the formula used is DMEM+20%FBS+10%DMSO+50ug / ml antifreeze protein (extracted from winter wheat plant)

[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 5×106 / ml~1×107 / 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:...

Embodiment 2

[0052] (1) Cell cryopreservation:

[0053] Freezing solution formula:

[0054]

[0055] 1. Prepare cell freezing culture medium, the formula used is DMEM+20%FBS+10%DMSO+50ug / ml antifreeze protein (extracted from winter wheat plant)

[0056] 2. Take the cells in the logarithmic growth phase, remove the old culture medium, and wash with PBS.

[0057] 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;

[0058] 4. Centrifuge at 1000rpm for 5min;

[0059] 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 5×106 / ml~1×107 / ml;

[0060] 6. Divide the cells into cryopreservation tubes, 1-1.5ml per tube;

[0061] 7. Mark the name of the cells, the time of freezing and the operator on the cryopreservation tube;

[0062] 8. Cryopreservatio...

Embodiment 3

[0070] (1) Cell cryopreservation:

[0071] Freezing solution formula:

[0072]

[0073] 1. Prepare cell freezing culture medium, the formula used is DMEM+20%FBS+10%DMSO+50ug / ml antifreeze protein (extracted from winter wheat plant)

[0074] 2. Take the cells in the logarithmic growth phase, remove the old culture medium, and wash with PBS.

[0075] 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;

[0076] 4. Centrifuge at 1000rpm for 5min;

[0077] 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 5×106 / ml~1×107 / ml;

[0078] 6. Divide the cells into cryopreservation tubes, 1-1.5ml per tube;

[0079] 7. Mark the name of the cells, the time of freezing and the operator on the cryopreservation tube;

[0080] 8. Cryopreservatio...

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PUM

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Abstract

The invention relates to the cell field, in particular to cell cryopreservation composition and an application thereof. Antifreeze protein is used in a cell cryopreservation liquid, so that the survival rate of cells is increased significantly. The antifreeze protein is added to the antifreeze composition, the survival rate can be significantly increased to 80% or above, the composition has greatsignificance in preserving rare cells such as stem cells, nerve cells and the like, meanwhile, the cell culture cost is also reduced after cell thawing, and the cell culture cycle is shortened.

Description

technical field [0001] The invention relates to the field of cells, in particular to a cell cryopreservation composition and application thereof. 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] 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 cells can also be purchased, donated, exchanged and shipped in the fo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 许澎黄巧
Owner 湖南丰晖生物科技有限公司
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