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Method for improving survival rate of human diploid cells

A technology for human diploid cells and survival rate, applied in the field of improving the survival rate of human diploid cells, can solve the problems of low survival rate of cryopreserved human diploid cells after resuscitation and recovery, and achieve the effect of improving the survival rate

Inactive Publication Date: 2015-03-25
CHENGDU KANGHUA BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to solve the defects of the prior art in the recovery of cryopreserved human diploid cells and the low survival rate

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0009] Preparation of cryopreservation solution

[0010] Take 1L of normal saline or TC199 serum-free medium, add 30ml of human serum albumin, 50ml of dimethyl sulfoxide, 5g of sodium carbonate, 20g of hydroxyethyl starch, 50g of serine, and 80g of dimethylacetamide; mix well stand-by.

Embodiment 1

[0012] After digesting the cells with trypsin solution, discard the trypsin in the digestion solution, add cell culture solution containing 10% calf serum to suspend the cells, centrifuge at 800-1000r / min for 5min, and discard the supernatant;

[0013] Resuspend the cell pellet with cryopreservation medium to a cell density of 2×10 6 pcs / ml~10 7 pcs / ml; packaged in a cryopreservation tube, let it stand for 15 hours at minus 80°C, and then quickly put it in liquid nitrogen for storage;

[0014] The solvent of the cryopreservation solution is physiological saline; each milliliter of the solvent contains 3% of human serum albumin, 5% of dimethyl sulfoxide, 0.5% of sodium carbonate, 2% of hydroxyethyl starch, 5% of serine, and dimethylacetamide 6%.

Embodiment 2

[0016] After digesting the cells with trypsin solution, discard the trypsin in the digestion solution, add cell culture solution containing 10% calf serum to suspend the cells, centrifuge at 800-1000r / min for 5min, and discard the supernatant;

[0017] Resuspend the cell pellet with cryopreservation medium to a cell density of 2×10 6 pcs / ml~10 7 pcs / ml; packaged in a cryopreservation tube, let it stand for 15 hours at minus 80°C, and then quickly put it in liquid nitrogen for storage;

[0018] The solvent of the freezing solution is TC199 serum-free medium; each milliliter of solvent contains 3% human serum albumin, 5% dimethyl sulfoxide, 0.5% sodium carbonate, 2% hydroxyethyl starch, 5% serine, dimethyl Acetamide 8%.

[0019] Control group 1

[0020] After digesting the cells with trypsin solution, discard the trypsin in the digestion solution, add cell culture solution containing 10% calf serum to suspend the cells, centrifuge at 800-1000r / min for 5min, and discard the su...

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PUM

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Abstract

The invention discloses a method for improving a survival rate of human diploid cells. The method comprises the following steps: digesting the cells by using a trypsin solution, centrifuging, and removing supernatant; suspending the cells by using a frozen stock solution, wherein a solvent of the frozen stock solution is normal saline or TC199 serum-free medium, and each milliliter of solvent contains 3 percent of human albumin, 5 percent of dimethyl sulfoxide, 0.5 percent of sodium carbonate, 2 percent of hydroxyethyl starch, 5 percent of serine and 6-10 percent of dimethylacetamide. According to the preservation method provided by the invention, the survival rate of the human diploid cells can be improved.

Description

technical field [0001] The invention relates to a method for cell preservation, in particular to a method for improving the survival rate of human diploid cells. Background technique [0002] The human diploid cell line is derived from normal human fetal tissue, and is mainly used for culturing viruses to prepare vaccines, etc. Human diploid cells need to be preserved after culture, and thawed and recovered when needed. It is well known that when cells are frozen and revived in liquid ammonia, it is impossible for all cells to survive. There are many factors that affect the survival rate of cell resuscitation. The existing research technology shows that the cryopreservation solution, cryopreservation method, and resuscitation method all have a direct impact on the survival rate of cell resuscitation. Contents of the invention [0003] The purpose of the present invention is to solve the defect of the prior art in the recovery and recovery of cryopreserved human diploid c...

Claims

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Application Information

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IPC IPC(8): A01N1/02
Inventor 侯文礼周蓉
Owner CHENGDU KANGHUA BIOLOGICAL PROD
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