In vitro culture of bone marrow-derived mesenchymal stem cells in a chemically-defined serum-free medium.
- Author:
Wei WU
1
;
Yan ZHOU
;
Wensong TAN
Author Information
1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Bone Marrow Cells;
cytology;
Cell Culture Techniques;
Cell Differentiation;
Cells, Cultured;
Culture Media, Serum-Free;
Mesenchymal Stromal Cells;
cytology;
Rabbits
- From:
Chinese Journal of Biotechnology
2009;25(1):121-128
- CountryChina
- Language:Chinese
-
Abstract:
This study is aimed to design a chemically-defined serum free medium (CDSFM) to support in vitro culture of bone marrow-derived mesenchymal stem cells (BM-MSCs). BM-MSCs were isolated from the femoral bones of one-month-old New Zealand Rabbits with density gradient centrifugation. We compared the proliferation capability, cell cycle, colony-forming efficiency, osteogenic and adipogenic differentiation capabilities of BM-MSCs cultured in CDSFM with those cultured in serum-containing medium (SCM). After 10 days culture, BM-MSCs were expanded by 50 folds in CDSFM, while only 40 folds in SCM. EGF, bFGF and hy-drocortisone were the most important additives and significantly stimulated BM-MSCs proliferation. The percentage of cells at G0-G1 cell cycle was 80.31% +/- 0.58% after CDSFM culture, with no significant difference (P>0.05) compared to 75.24% +/- 4.05% for SCM culture. However, the cloning efficiency of BM-MSCs cultured in CDSFM was significantly lower than that in SCM (P<0.01). The expanded BM-MSCs in CDSFM preserved differentiation potentials into mesenchymal lineages in vitro, including adipocytes and osteoblasts. We have designed a chemically-defined serum free medium that could support in vitro proliferation and maintain the properties of BM-MSCs as stem cells, which could be applied to cell-based therapy and biomedical research.
