A cell cryopreservation liquid for human umbilical cord mesenchymal stem cells

A technology of stem cells and human umbilical cord, applied in the field of cell cryopreservation of human umbilical cord mesenchymal stem cells, can solve the problems of pollution, reduce antibiotic content, increase cell survival rate, etc.

Inactive Publication Date: 2018-11-16
QINGDAO RESTORE BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve the existing technical task of the present invention is to solve the deficiencies of the existing cryopreservation conditions, provide a cryopreservation solution of human umbilical cord mesenchymal stem cells, and solve the problem of animal fetal bovine serum in the conventional cryopreservation solution. Source components and pathogen pollution, reduce cell death and maintain stem cell characteristics, added streptomycin sulfate, can reduce antibiotic content, reduce the toxicity of antibiotics to human umbilical cord mesenchymal stem cells, increase cell survival rate, and promote cell growth

Method used

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  • A cell cryopreservation liquid for human umbilical cord mesenchymal stem cells
  • A cell cryopreservation liquid for human umbilical cord mesenchymal stem cells

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Experimental program
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Effect test

Embodiment 1

[0026] A cell cryopreservation solution for human umbilical cord mesenchymal stem cells, composed of serum-free culture medium, serum substitute, dimethyl sulfoxide, streptomycin sulfate and additives, wherein the percentage of ingredients is: serum-free culture medium: 41.8 %, serum substitute: 50%, dimethylsulfoxide: 8%, 200ug / ml streptomycin sulfate: 0.1%, additive: 0.1%.

[0027] The serum-free medium contains the following components added to the basal culture medium, and the final concentrations of each component are: human serum albumin 1-2 g / L, transferrin 5-10 mg / L, fibronectin 2-2 8mg / L, laminin 1~4mg / L, Fe(NO3)3 9H2O 50g / L, FeSO4 7H2O4 17g / L, estradiol 1~3μg / L, testosterone 2~5μg / L, progesterone 1~3μg / L, dexamethasone 39.25~117.74μg / L, insulin 5~10mg / L, riboflavin 376.36mg / L, coenzyme A80.96~242.87mg / L, butyldiamine 4.41~6.17mg / L, taurine Acid 1~2mg / L, aminoethanol 0.61~1.85mg / L, pyruvic acid 8.81~26.42mg / L, sodium selenate 3.78~7.56μg / L, L-glutamine 292.3~584.6mg...

Embodiment 2

[0038]A cell cryopreservation solution for human umbilical cord mesenchymal stem cells, composed of serum-free culture medium, serum substitute, dimethyl sulfoxide, streptomycin sulfate and additives, wherein the percentage of ingredients is: serum-free culture medium: 46.8 %, serum substitute: 45%, dimethylsulfoxide: 8%, 200ug / ml streptomycin sulfate: 0.1%, additive: 0.1%.

[0039] The serum-free medium contains the following components added to the basal culture medium, and the final concentrations of each component are: human serum albumin 1-2 g / L, transferrin 5-10 mg / L, fibronectin 2-2 8mg / L, laminin 1~4mg / L, Fe(NO3)3 9H2O 50g / L, FeSO4 7H2O4 17g / L, estradiol 1~3μg / L, testosterone 2~5μg / L, progesterone 1~3μg / L, dexamethasone 39.25~117.74μg / L, insulin 5~10mg / L, riboflavin 376.36mg / L, coenzyme A80.96~242.87mg / L, butyldiamine 4.41~6.17mg / L, taurine Acid 1~2mg / L, aminoethanol 0.61~1.85mg / L, pyruvic acid 8.81~26.42mg / L, sodium selenate 3.78~7.56μg / L, L-glutamine 292.3~584.6mg / ...

Embodiment 3

[0050] A cell cryopreservation solution for human umbilical cord mesenchymal stem cells, composed of serum-free culture medium, serum substitute, dimethyl sulfoxide, streptomycin sulfate and additives, wherein the percentage of ingredients is: serum-free culture medium: 51.8 %, serum substitute: 40%, dimethylsulfoxide: 8%, 200ug / ml streptomycin sulfate: 0.1%, additive: 0.1%.

[0051] The serum-free medium contains the following components added to the basal culture medium, and the final concentrations of each component are: human serum albumin 1-2 g / L, transferrin 5-10 mg / L, fibronectin 2-2 8mg / L, laminin 1~4mg / L, Fe(NO3)3 9H2O 50g / L, FeSO4 7H2O4 17g / L, estradiol 1~3μg / L, testosterone 2~5μg / L, progesterone 1~3μg / L, dexamethasone 39.25~117.74μg / L, insulin 5~10mg / L, riboflavin 376.36mg / L, coenzyme A80.96~242.87mg / L, butyldiamine 4.41~6.17mg / L, taurine Acid 1~2mg / L, aminoethanol 0.61~1.85mg / L, pyruvic acid 8.81~26.42mg / L, sodium selenate 3.78~7.56μg / L, L-glutamine 292.3~584.6mg...

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Abstract

A cell cryopreservation liquid for human umbilical cord mesenchymal stem cells is disclosed. Components of the liquid include 41.8-51.8% of a serum-free culture solution, 40-50% of a serum substitute,8% of dimethyl sulfoxide, 0.1% of streptomycin sulfate having a concentration of 200 [mu]g / mL and 0.1% of an additive. A cryopreservation method utilizing the cell cryopreservation liquid for the human umbilical cord mesenchymal stem cells is further provided. The method includes centrifuging suspension cells after subculture, then putting the cells into a centrifuge tube, adding the cryopreservation liquid into the centrifuge tube containing the cells, transferring the mixture to a cryopreservation tube, putting the cryopreservation tube requiring cryopreservation into a programmed cooling box containing isopropanol, allowing the cryopreservation tube to stay overnight at -80 DEG C and then transferring the cryopreservation tube to a liquid nitrogen tank the next day. A problem that fetal calf serum in conventional cryopreservation liquids brings about animal-sourced components and pathogen contamination can be solved. The added streptomycin sulfate can reduce the content of antibiotics, reducing human umbilical cord mesenchymal stem cell poisoning caused by antibiotics, reducing cell death and maintaining characteristics of the stem cells.

Description

technical field [0001] The invention relates to cell culture technology, in particular to a cell cryopreservation solution of human umbilical cord mesenchymal stem cells. Background technique [0002] Human umbilical cord mesenchymal stem cells refer to a kind of pluripotent stem cells isolated from umbilical cord. Compared with other sources of mesenchymal stem cells, they have obvious advantages in clinical application, such as easy to obtain materials, easy to collect, cryopreservation and relatively pure, etc. High differentiation potential, can be differentiated in multiple directions, such as osteoblasts, chondrocytes, hepatocytes, insulin-like cells, nerve cells and cardiomyocytes, etc., and there are no ethical issues. At present, human umbilical cord mesenchymal stem cells have been applied in the clinical field and achieved encouraging therapeutic effects. [0003] The seed bank of human umbilical cord mesenchymal stem cells has been widely used clinically, but th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 戚凤春宫丽张炳强章宏金铎于淼于露
Owner QINGDAO RESTORE BIOTECHNOLOGY CO LTD
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