Meuleman Nathalie, Tondreau Tatiana, Delforge Alain, Dejeneffe Marielle, Massy Martine, Libertalis Mark, Bron Dominique, Lagneaux Laurence
Department of Experimental Hematology, Institut Jules Bordet, Centre des Tumeurs de l'Université Libre de Bruxelles, Brussels, Belgium.
Eur J Haematol. 2006 Apr;76(4):309-16. doi: 10.1111/j.1600-0609.2005.00611.x.
The expansion of mesenchymal stem cells (MSCs) strongly depends on the culture conditions and requires medium supplemented with 10-20% fetal calf serum (FCS) to generate relevant numbers of cells. However, the presence of FCS is a major obstacle for their clinical use. Therefore, we have evaluated the capacity of expansion of MSC in a commercial serum-free medium (UC) supplemented with a serum substitute (ULTROSER) in comparison with a classical medium alpha-MEM containing 15% FBS. Bone marrow-mononuclear cells collected from 12 volunteer healthy donors were expanded in two different culture media. MSCs isolated in the both media were morphologically similar and expressed identical phenotypic markers. After the primoculture (P0) and one passage, we obtained significantly more MSC and CFU-F progenitors in UC medium than in alphaMEM. Their multipotentiality was preserved during culture, as well as their capacity to support haematopoiesis. In conclusion, our observations strongly suggest that UC is an optimal medium for ex vivo expansion of MSC: it allows a better cell expansion, preserves cell multipotentiality, reduces the culture period and contains low concentration of serum substitute. This medium seems suitable for clinical scale expansion of MSC.
间充质干细胞（MSCs）的扩增强烈依赖于培养条件，需要添加10 - 20%胎牛血清（FCS）的培养基才能产生足够数量的细胞。然而，FCS的存在是其临床应用的主要障碍。因此，我们评估了与含有15%胎牛血清的经典α - MEM培养基相比，在添加了血清替代物（ULTROSER）的商业无血清培养基（UC）中MSCs的扩增能力。从12名健康志愿者供体采集的骨髓单个核细胞在两种不同的培养基中进行扩增。在两种培养基中分离得到的MSCs在形态上相似且表达相同的表型标志物。在原代培养（P0）和传代一次后，我们在UC培养基中获得的MSC和CFU - F祖细胞比在α - MEM中显著更多。它们的多能性在培养过程中得以保留，支持造血的能力也得以保留。总之，我们的观察结果强烈表明UC是用于MSCs体外扩增的最佳培养基：它能实现更好的细胞扩增，保留细胞多能性，缩短培养周期且血清替代物浓度低。这种培养基似乎适合MSCs的临床规模扩增。
