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Culture medium for amplification of human umbilical cord mesenchymal stem cells and culture method

A technique for stromal stem cells and culturing methods, which is applied in the field of culture medium for expansion of human umbilical cord mesenchymal stem cells, can solve the problems of poor activity, low content of cytokines in the medium, slow cell growth, etc. Increased proliferation efficiency and clear ingredients

Active Publication Date: 2021-07-23
山西北科生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many unknown components in the serum, and may even include some unknown allergens and pathogens, which cannot guarantee the safety of the cultured cells, which hinders the clinical application of cells
At present, the serum-free medium used for umbilical cord mesenchymal stem cells is not good for cell culture, and there are problems such as slow cell growth and poor activity, and low content of cytokines in the medium, which affects the promotion of umbilical cord mesenchymal stem cell biological products. application

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A medium for expansion of human umbilical cord mesenchymal stem cells, which is composed of DMEM / F12 medium and the following components added to the above medium: glutathione, α-tocopherol, cineole, estradiol valerate Alcohol, acetazolamide, sodium selenite, vitamin C, insulin; based on the final concentration, the dosage of each component is: glutathione 14μg / mL, α-tocopherol 2μg / mL, eucalyptol 7.5ng / mL mL, estradiol valerate 8ng / mL, acetazolamide 4.5ng / mL, sodium selenite 7μg / mL, vitamin C 10μg / mL, insulin 13μg / mL.

[0029] A method for culturing human umbilical cord mesenchymal stem cells, comprising the following steps:

[0030] (1) Take DMEM / F12 medium, add glutathione, α-tocopherol, cineole, estradiol valerate, acetazolamide, sodium selenite, vitamin C, insulin, and mix well;

[0031] (2) Inoculate the human umbilical cord mesenchymal stem cells to be cultured in the medium of step (1), and the density of the human umbilical cord mesenchymal stem cells inoculate...

Embodiment 2

[0033] A medium for expansion of human umbilical cord mesenchymal stem cells, which is composed of DMEM / F12 medium and the following components added to the above medium: glutathione, α-tocopherol, cineole, estradiol valerate Alcohol, acetazolamide, sodium selenite, vitamin C, insulin; in terms of final concentration, the dosage of each component is: glutathione 12μg / mL, α-tocopherol 1μg / mL, eucalyptol 5.5ng / mL mL, estradiol valerate 5 ng / mL, acetazolamide 3.5 ng / mL, sodium selenite 5 μg / mL, vitamin C 8 μg / mL, insulin 10 μg / mL.

[0034] A method for culturing human umbilical cord mesenchymal stem cells, comprising the following steps:

[0035] (1) Take DMEM / F12 medium, add glutathione, α-tocopherol, cineole, estradiol valerate, acetazolamide, sodium selenite, vitamin C, insulin, and mix well;

[0036] (2) Inoculate the human umbilical cord mesenchymal stem cells to be cultured in the medium of step (1), and the density of the human umbilical cord mesenchymal stem cells inocul...

Embodiment 3

[0038] A medium for expansion of human umbilical cord mesenchymal stem cells, which is composed of DMEM / F12 medium and the following components added to the above medium: glutathione, α-tocopherol, cineole, estradiol valerate Alcohol, acetazolamide, sodium selenite, vitamin C, insulin; in terms of final concentration, the dosage of each component is: glutathione 16μg / mL, α-tocopherol 3μg / mL, cineole 8.5ng / mL mL, estradiol valerate 10 ng / mL, acetazolamide 5 ng / mL, sodium selenite 10 μg / mL, vitamin C 11 μg / mL, insulin 15 μg / mL.

[0039] A method for culturing human umbilical cord mesenchymal stem cells, comprising the following steps:

[0040] (1) Take DMEM / F12 medium, add glutathione, α-tocopherol, cineole, estradiol valerate, acetazolamide, sodium selenite, vitamin C, insulin, and mix well;

[0041] (2) Inoculate the human umbilical cord mesenchymal stem cells to be cultured in the medium of step (1), and the density of the human umbilical cord mesenchymal stem cells inoculat...

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Abstract

The invention discloses a culture medium for amplification of human umbilical cord mesenchymal stem cells. The culture medium is prepared from a basic culture medium and the following components added into the basic culture medium of glutathione, alpha-tocopherol, eudesmin, estradiol valerate, acetazolamide, sodium selenite, vitamin C and insulin. Exogenous serum is not added into the culture medium provided by the invention, the components are clear, and the components have a synergistic effect, so that the proliferation of the umbilical cord mesenchymal stem cells and the secretion of cell factors are promoted. The added ingredients such as eudesmin, estradiol valerate, acetazolamide and the like not only avoid adding serum into the culture medium to form ingredients with potential safety hazards, but also can improve the proliferation efficiency of cells, a large number of umbilical cord mesenchymal stem cells can be obtained within a short time, and the content of cell factors in the culture solution is increased. The invention also provides a culture method of the human umbilical cord mesenchymal stem cells, which adopts the culture medium provided by the invention, and is safe, efficient, simple and convenient in process and easy to operate.

Description

technical field [0001] The invention relates to the field of stem cells, in particular to a culture medium and a culture method for the expansion of human umbilical cord mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (MSCs) are pluripotent stem cells derived from mesoderm with high self-renewal ability and multi-directional differentiation potential. At present, mesenchymal stem cells are mainly extracted from umbilical cord, bone marrow, adipose tissue, dental pulp, and placenta. Mesenchymal stem cells still have multi-directional differentiation potential after continuous subculture and cryopreservation, which provides a new way for the clinical treatment of various diseases, such as nervous system diseases, kidney diseases, autoimmune diseases, and malignant tumors. [0003] Umbilical cord mesenchymal stem cells refer to a kind of multifunctional stem cells existing in the umbilical cord tissue of newborns. seed cells. In addition, umbilic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0668C12N2501/999C12N2500/38C12N2501/33C12N2501/392C12N2501/60C12N2501/70C12N2500/12
Inventor 董润润陈玲王建声
Owner 山西北科生物科技有限公司
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