Wu Wei, Zhou Yan, Tan Wensong
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
Sheng Wu Gong Cheng Xue Bao. 2009 Jan;25(1):121-8.
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.
本研究旨在设计一种化学成分明确的无血清培养基（CDSFM），以支持骨髓间充质干细胞（BM-MSCs）的体外培养。通过密度梯度离心法从1月龄新西兰兔的股骨中分离出BM-MSCs。我们比较了在CDSFM中培养的BM-MSCs与在含血清培养基（SCM）中培养的BM-MSCs的增殖能力、细胞周期、集落形成效率、成骨和成脂分化能力。培养10天后，BM-MSCs在CDSFM中扩增了50倍，而在SCM中仅扩增了40倍。表皮生长因子（EGF）、碱性成纤维细胞生长因子（bFGF）和氢化可的松是最重要的添加剂，能显著刺激BM-MSCs的增殖。CDSFM培养后处于G0-G1细胞周期的细胞百分比为80.31%±0.58%，与SCM培养的75.24%±4.05%相比，无显著差异（P>0.05）。然而，在CDSFM中培养的BM-MSCs的克隆效率显著低于在SCM中的克隆效率（P<0.01）。在CDSFM中扩增的BM-MSCs在体外保留了向间充质谱系分化的潜能，包括脂肪细胞和成骨细胞。我们设计了一种化学成分明确的无血清培养基，可支持BM-MSCs的体外增殖并维持其作为干细胞的特性，可应用于基于细胞的治疗和生物医学研究。
