Serum-free culture medium for mesenchymal stem cells
A serum-free medium and mesenchymal stem cell technology, applied in the field of mesenchymal stem cell serum-free medium, can solve the problems of affecting the growth of keratinocytes, accelerating keratinocyte differentiation, and fibroblast pollution, so as to prevent the occurrence and development of cancer , reduce blood viscosity, inhibit vasodilation
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Embodiment 1
[0038] Embodiment one : Adipose-derived mesenchymal stem cells are subcultured in culture medium;
[0039] Medium 1: serum-free medium for mesenchymal stem cells provided by the present invention;
[0040] Medium 2: L-DMEM+10%FBS (Fetal Bovine Serum);
[0041] Medium 3: STEMPRO MSC SFM serum-free medium.
[0042] The above-mentioned L-DMEM is provided by Gibco Company, the article number is 11885-084.
[0043] The above-mentioned STEMPRO MSC SFM serum-free medium is provided by Gibco Company, the product number is A10332-01.
[0044] Steps:
[0045] 1.1 Dilute adipose-derived mesenchymal stem cells at 10,000 cells / cm 2 The density was planted into three T75 cell culture flasks, and medium 1, medium 2, and medium 3 were added respectively. After the medium and the adipose-derived mesenchymal stem cell suspension were fully mixed, the three cell culture flasks were put into to 37°C, CO 2 The concentration was 5% in the cell culture incubator, and thereafter every 3 days a...
Embodiment 2
[0052] Embodiment two : Umbilical cord mesenchymal stem cells are subcultured in culture medium;
[0053] Medium 1: serum-free medium for mesenchymal stem cells provided by the present invention;
[0054] Medium 2: L-DMEM+10%FBS (Fetal Bovine Serum);
[0055] Medium 3: STEMPRO MSC SFM serum-free medium.
[0056] The above-mentioned L-DMEM is provided by Gibco Company, the article number is 11885-084.
[0057] The above-mentioned STEMPRO MSC SFM serum-free medium is provided by Gibco Company, the product number is A10332-01.
[0058] Steps:
[0059] 2.1. The umbilical cord mesenchymal stem cells were divided into 10000 cells / cm 2 The density was planted into three T75 cell culture bottles, and medium 1, medium 2, and medium 3 were added respectively. After the medium and the umbilical cord mesenchymal stem cell suspension were fully mixed, the three cell culture bottles were put into to 37°C, CO 2 Cell culture was carried out in a cell incubator with a concentration of ...
Embodiment 3
[0066] Embodiment three: Biological characteristics of umbilical cord-derived mesenchymal stem cells and adipose-derived mesenchymal stem cells subcultured in medium;
[0067] Medium: serum-free medium for mesenchymal stem cells provided by the present invention.
[0068] Steps:
[0069] 3.1 The umbilical cord mesenchymal stem cells and adipose-derived mesenchymal stem cells were divided into 10000 cells / cm 2 The densities were respectively planted in 2 T175 cell culture flasks, added to the serum-free medium of mesenchymal stem cells provided by the present invention for subculture, and each type of stem cells were subcultured 5 times and 10 times respectively;
[0070] 3.2 Harvest the cells in the cell culture flask, digest with 0.25% trypsin for 2 min, then blow off the mesenchymal stem cells and terminate the digestion with medium;
[0071] 3.3 Centrifuge the umbilical cord-derived mesenchymal stem cells and adipose-derived mesenchymal stem cells obtained in step 3.2 a...
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