Jung Jooyoung, Moon Noory, Ahn Ji-Yeon, Oh Eun-Jee, Kim Myungshin, Cho Chul-Soo, Shin Jong-Chul, Oh Il-Hoan
Department of Cellular Medicine, Catholic High-Performance Cell Therapy Center, The Catholic University of Korea, Seoul, Korea.
Stem Cells Dev. 2009 May;18(4):559-71. doi: 10.1089/scd.2008.0105.
Recent clinical trials using ex vivo expanded mesenchymal stromal cells (MSCs) have raised interest in the safety and function of cultured MSCs. Here, to assess the feasibility of using allogenic human umbilical cord blood serum (CBS) for humanized clinical-grade expansion of MSCs, we characterized MSCs expanded in CBS and compared them to MSCs expanded in fetal bovine serum (FBS). MSCs in CBS exhibited a higher preservation of colony-forming cells and an accelerated expansion over serial passages with increased Oct-4 expression compared to those cultured in FBS. Notably, CBS-expanded MSCs exhibited a unique differentiation potential characterized by a shift from adipogenic to osteogenic differentiation. The differentiation shift was associated with enhanced basal and Runx2-mediated transcriptional activation of the osteocalcin promoter, as well as increased accumulation of beta-catenin and the yes-associated protein (YAP) which was independent of changes in TAZ (transcriptional co-activator with PDZ-binding motif) levels. Interestingly, the phenotypes were reversed when the FBS and CBS media were switched, suggesting the unique stimulatory effects of CBS rather than the selection of heterogeneous MSC subpopulations. The distinct regulatory effects of CBS on MSC included selective activation of platelet-derived growth factor and epidermal growth factor signals in MSCs, but not in FBS. Taken together, these results provide insight into the dynamic regulation of MSCs during ex vivo culture and show that the ex vivo culture of MSCs in allogenic human CBS provides a novel tool for the accelerated expansion of a population of MSCs that exhibit a higher self-renewal and an enhanced osteogenic potential.
近期使用体外扩增间充质基质细胞（MSC）的临床试验引发了人们对培养的MSC安全性和功能的关注。在此，为评估使用同种异体人脐带血血清（CBS）进行人源化临床级MSC扩增的可行性，我们对在CBS中扩增的MSC进行了表征，并将其与在胎牛血清（FBS）中扩增的MSC进行比较。与在FBS中培养的MSC相比，CBS中的MSC在连续传代过程中表现出更高的集落形成细胞保留率和加速扩增，同时Oct-4表达增加。值得注意的是，CBS扩增的MSC表现出独特的分化潜能，其特征是从脂肪生成向成骨分化的转变。这种分化转变与骨钙素启动子的基础和Runx2介导的转录激活增强有关，以及β-连环蛋白和Yes相关蛋白（YAP）的积累增加，这与具有PDZ结合基序的转录共激活因子（TAZ）水平的变化无关。有趣的是，当FBS和CBS培养基切换时，表型会逆转，这表明CBS具有独特的刺激作用，而不是选择了异质性的MSC亚群。CBS对MSC的独特调节作用包括选择性激活MSC中的血小板衍生生长因子和表皮生长因子信号，但在FBS中则不然。综上所述，这些结果为体外培养过程中MSC的动态调节提供了见解，并表明在同种异体人CBS中体外培养MSC为加速扩增具有更高自我更新能力和增强成骨潜能的MSC群体提供了一种新工具。
