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Cell cryopreservation liquid containing antifreeze proteins

A technology of antifreeze protein and cryopreservation solution, which is applied in the field of cell cryopreservation solution containing antifreeze protein, which can solve the problems of human body and cytotoxicity

Inactive Publication Date: 2017-11-10
昆明医科大学第二附属医院
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, DMSO is toxic to the human body and cells, and can cause morphological differentiation of cells

Method used

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  • Cell cryopreservation liquid containing antifreeze proteins
  • Cell cryopreservation liquid containing antifreeze proteins

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0025] (1) Preparation of cryopreservation solution:

[0026] Weigh AFP134 (final concentration 0.01mg / ml) 1mg, calf serum (final concentration 90%) 90ml, sucrose (final concentration 0.5M / L) 17g, sodium citrate (final concentration 0.05M / L) 1.47g

[0027] Add the above substances into 90ml sterilized double distilled water, and dilute to 100ml

[0028] (2) Cell cryopreservation

[0029] 1. Take the cells in the logarithmic growth phase, remove the old culture medium, and wash with PBS.

[0030] 2. Remove the PBS, add an appropriate amount of trypsin (covering the surface of the culture dish) to digest the cells growing in the monolayer;

[0031] 3. Centrifuge at 1000rpm for 5min;

[0032] 4. Remove trypsin, add an appropriate amount of prepared cryopreservation solution, blow gently with a pipette to make the cells uniform, count, and adjust the final density of cells in the cryopreservation solution to 5×10 6 / ml;

[0033] 5. Divide the cells into cryopreservation tubes...

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Abstract

The invention relates to the field of cell freezing liquids, and concretely relates to a cell cryopreservation liquid containing antifreeze proteins. AFP134 proteins are added to the cell cryopreservation liquid in order to effectively improve the cell recovery rate; and the cell cryopreservation liquid contains no DMSO, so the influences of the DMSO on the cytotoxicity are reduced.

Description

technical field [0001] The invention relates to the field of cell cryopreservation solution, in particular to a cell cryopreservation solution containing antifreeze protein. Background technique [0002] Cell cryopreservation is one of the main methods of cell preservation. Using cryopreservation technology to store cells in -196°C liquid nitrogen at ultra-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. [0003] During cryopreservation of cells at ultra-low temperatures (minus 200 degrees), it is necessary to use cryoprotectants to prevent damage caused by intracellular liquid ice crystal formation, osmotic pressure changes, and cell structure diso...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 王燕奎翔易晓佳
Owner 昆明医科大学第二附属医院
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