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Culture medium and culture method for mesenchymal stem cells

A technology of mesenchymal stem cells and culture medium, applied in the field of cell biology, can solve the problems of lack of culture medium and culture methods, and achieve the effects of improving the efficiency of osteogenic differentiation, shortening the time of calcium nodules, and increasing the expression

Pending Publication Date: 2020-11-17
海南济民博鳌国际医院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is still a lack of simple and efficient culture media and culture methods for the osteogenic differentiation of mesenchymal stem cells

Method used

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  • Culture medium and culture method for mesenchymal stem cells
  • Culture medium and culture method for mesenchymal stem cells

Examples

Experimental program
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Effect test

Embodiment 1

[0028] A medium for adipose-derived mesenchymal stem cells, including the following components: luteolin 30 μg / mL, fisetin 10 μg / mL, emodin 15 μg / mL, 1,2,3,6-tetra-O- Galloyl-β-D-glucose 10μg / mL, penicillin 20U / mL, streptomycin 50mg / mL, L-ascorbic acid 0.04mM, L-glutamine 3mM, epidermal growth factor 16ng / mL, sodium pyruvate 1mM, and The volume is DMEM low-glucose medium.

Embodiment 2

[0030] A medium for adipose-derived mesenchymal stem cells, including the following components: luteolin 15 μg / mL, fisetin 22 μg / mL, emodin 30 μg / mL, 1,2,3,6-tetra-O- Galloyl-β-D-glucose 5μg / mL, penicillin 30U / mL, streptomycin 60mg / mL, L-ascorbic acid 0.02mM, L-glutamine 6mM, epidermal growth factor 10ng / mL, sodium pyruvate 2mM, The volume is DMEM low-glucose medium.

[0031] The method for culturing adipose-derived mesenchymal stem cells using the culture medium of Example 1 and Example 2: Take mesenchymal stem cells and culture them with culture medium, pH 7.0-7.3; change the medium every 2 days.

Embodiment 3

[0033] The method for culturing adipose-derived mesenchymal stem cells using the culture medium of the present invention:

[0034] (1) Take mesenchymal stem cells and culture them with medium for 2 days. The medium is: luteolin 23 μg / mL, 1,2,3,6-tetra-O-galloyl-β-D-glucose 8 μg / mL, penicillin 30U / mL, streptomycin 60mg / mL, L-ascorbic acid 0.04mM, L-glutamine 6mM, epidermal growth factor 16ng / mL, sodium pyruvate 2mM, the balance is DMEM low-sugar medium, pH7. 0~7.3;

[0035] (2) Replacement of the culture medium, the culture medium is 20 μg / mL of fisetin and 25 μg / mL of emodin added to the medium of step (1), pH7.0~7.3;

[0036] (3) Afterwards, the medium was replaced every 2 days, and the composition of the medium was the same as in step (2).

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Abstract

The invention discloses a culture medium and a culture method for mesenchymal stem cells. The culture medium comprises luteolin, fisetin, emodin, 1, 2, 3, 6-tetra-O-galloyl-beta-D-glucose, penicillin,streptomycin, L-ascorbic acid, L-glutamine, epidermal growth factors and the like. By using the culture medium, the time of obvious calcium nodules is effectively shortened by at least 5 days, the expression of osteopontin mRNA is significantly increased at the same culture time, and the osteogenic differentiation efficiency of adipose-derived mesenchymal stem cells is improved.

Description

technical field [0001] The invention relates to the technical field of cell biology, in particular to a culture medium for mesenchymal stem cells and a preparation method thereof. Background technique [0002] Mesenchymal stem cells are a type of adult stem cells that exist in a variety of tissues (such as bone marrow, umbilical cord blood and umbilical cord tissue, placental tissue, adipose tissue, etc.), have multi-directional differentiation potential, non-hematopoietic stem cells, and can differentiate into Fat cells, osteoblasts, chondrocytes, nerve cells, cardiomyocytes, endothelial cells, etc. Mesenchymal stem cells, as the source of cells for cell therapy of various diseases, have become a hot spot in the field of stem cell research. At present, there is still a lack of simple and efficient culture media and culture methods for the osteogenic differentiation of mesenchymal stem cells. Contents of the invention [0003] In view of the deficiencies in the prior art...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0662C12N5/0667C12N2500/38C12N2500/32C12N2500/30C12N2501/11C12N2501/06C12N2501/999Y02A50/30
Inventor 王忠伟李巍符春娱
Owner 海南济民博鳌国际医院有限公司
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