Institute of Experimental Medicine, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
Cytotherapy. 2009;11(7):874-85. doi: 10.3109/14653240903188947.
Human mesenchymal stromal cells (hMSC) are a promising tool for future clinical application, but their use requires rapid cell expansion in media suitable for clinical use. Therefore, we tested the influence of several culture media on colony formation, population doubling (PD) time, cell cycle and surface marker expression.
hMSC isolated from human bone marrow (BM) obtained from healthy donors were seeded and expanded in different culture media: alpha-minimum essential medium (MEM) supplemented with 2.5%, 5%, 10% or 20% fetal bovine serum (FBS), 5% or 10% human cord blood serum (hCBS), 5% or 10% human blood serum from AB adult donors (hABS), or mesenchymal stem cell growth medium (MSCGM). The number, diameter and total area of the colonies formed and PD time were determined, and the cell cycle and 16 surface markers were analyzed.
Colony-forming efficiency was best in alpha-MEM/hCBS and alpha-MEM/hABS, good in MSCGM and worst in alpha-MEM/FBS. The shortest PD time was achieved in media enriched with human sera or MSCGM, while the time was increased in alpha-MEM/FBS. The largest proliferating fraction was seen in MSCGM followed by media enriched with human sera; the fraction was smallest in alpha-MEM/FBS. Staining for CD34, CD45, CD235a and CD271 was negative, while that for CD29, CD44, CD73, CD90, CD105 and human leukocyte antigen (HLA)-A, -B, -C was positive in all media tested. Media with human serum did not adversely affect the differentiation potential of hMSC, and differentiation into osteoblasts was enhanced.
The choice of serum influences hMSC expansion and cell properties; alpha-MEM supplemented with hABS seems to be a promising candidate for clinical use.
人骨髓间充质基质细胞（hMSC）是未来临床应用的有前途的工具，但它们的使用需要在适合临床使用的培养基中快速扩增细胞。因此，我们测试了几种培养基对集落形成、倍增时间（PD）、细胞周期和表面标记物表达的影响。
从健康供体的人骨髓（BM）中分离的 hMSC 接种并在不同的培养基中扩增：补充有 2.5%、5%、10%或 20%胎牛血清（FBS）的α-最小必需培养基（MEM）、5%或 10%人脐血血清（hCBS）、5%或 10%来自 AB 成年供体的人血血清（hABS），或间充质干细胞生长培养基（MSCGM）。确定形成的集落的数量、直径和总面积以及 PD 时间，并分析细胞周期和 16 个表面标记物。
集落形成效率在 alpha-MEM/hCBS 和 alpha-MEM/hABS 中最佳，在 MSCGM 中良好，在 alpha-MEM/FBS 中最差。在富含人血清或 MSCGM 的培养基中实现最短的 PD 时间，而在 alpha-MEM/FBS 中时间增加。在 MSCGM 之后观察到最大的增殖分数，其次是富含人血清的培养基；在 alpha-MEM/FBS 中最小。CD34、CD45、CD235a 和 CD271 的染色为阴性，而 CD29、CD44、CD73、CD90、CD105 和人白细胞抗原（HLA）-A、-B、-C 的染色为阳性，所有测试的培养基均为阳性。含人血清的培养基不会对 hMSC 的分化潜能产生不利影响，并且增强了向成骨细胞的分化。
血清的选择会影响 hMSC 的扩增和细胞特性；补充有 hABS 的 alpha-MEM 似乎是临床应用的有前途的候选物。
