Chen H H, Decot V, Ouyang J P, Stoltz J F, Bensoussan D, de Isla N G
Faculty of Medicine, Nancy University, Vandoeuvre, France.
Biomed Mater Eng. 2009;19(4-5):301-9. doi: 10.3233/BME-2009-0595.
In the last years, there were many studies based on the use of human bone marrow mesenchymal stem cells (hMSCs) in cell therapy and tissue engineering. Although hMSCs can be easily obtained and expanded in culture, a large number of cells are often needed. The expansion of hMSCs depends on the culture conditions, such as media, cell density or culture flasks. Moreover, growth factors are often added to improve cell proliferation. In this study, we compared the effect of two culture media (DMEM and alpha-MEM), two culture flasks (75 or 25 cm2) and two different mononuclear cell seeding densities (1 x 10(4) or 5 x 10(4) MNC/cm2) on the isolation of hMSCs from bone marrow samples and analyzed if the isolation conditions affected the expansion of these cells in the first two passages. Experiments were performed without the addition of exogenous growth factors. Our results showed that alpha-MEM is the optimal culture medium for both, isolation and expansion of mesenchymal stem cells. Moreover, the cell seeding density of 50,000 MNC/cm2 in 25 cm2 culture flasks seems to be the best condition for the isolation step.
在过去几年中，有许多基于人骨髓间充质干细胞（hMSCs）用于细胞治疗和组织工程的研究。尽管hMSCs能够很容易地从培养物中获取并扩增，但通常需要大量细胞。hMSCs的扩增取决于培养条件，如培养基、细胞密度或培养瓶。此外，经常添加生长因子以促进细胞增殖。在本研究中，我们比较了两种培养基（DMEM和α-MEM）、两种培养瓶（75或25 cm²）以及两种不同的单核细胞接种密度（1×10⁴或5×10⁴个有核细胞/cm²）对从骨髓样本中分离hMSCs的影响，并分析了分离条件是否影响这些细胞在前两代中的扩增。实验在不添加外源性生长因子的情况下进行。我们的结果表明，α-MEM是用于间充质干细胞分离和扩增的最佳培养基。此外，在25 cm²培养瓶中接种密度为50,000个有核细胞/cm²似乎是分离步骤的最佳条件。
