CHA Stem Cell Institute, Department of Biomedical Science, CHA University College of Medicine, Seoul, 135-081, South Korea.
Stem Cell Rev Rep. 2010 Sep;6(3):425-37. doi: 10.1007/s12015-010-9158-x.
Human embryonic stem cell (hESC)-based assay systems and genetically modified hESCs are very useful tools for screening drugs that regulate stemness and differentiation and for studying the molecular mechanisms involved in hESC fate determination. For these types of studies, feeder cell-dependent cultures of hESCs are often problematic because the physiology of the feeder cells is perturbed by the drug treatments or genetic modifications, which potentially obscures research outcomes. In this study, we evaluated three commonly used feeder-free culture conditions to determine whether they supported the undifferentiated growth of hESCs and to determine whether the hESCs grown in these conditions displayed gene expression patterns that were similar to the expression patterns of feeder cell-dependent hESCs. Our results demonstrate that hESCs grown in the three feeder-free conditions expressed undifferentiation marker genes as strongly as hESCs that were grown in the feeder-dependent cultures. Furthermore, genome-wide gene expression profiles indicated that the gene expression patterns of hESCs that were grown under feeder-free or feeder-dependent culture conditions were highly similar. These results indicate that the feeder-free culture conditions support the undifferentiated growth of hESCs as effectively as the feeder-dependent culture conditions. Therefore, feeder-free culture conditions are potentially suitable for drug screening and for the genetic manipulation of hESCs in basic research.
基于人类胚胎干细胞(hESC)的检测系统和经过基因修饰的 hESC 是筛选调节干细胞特性和分化的药物以及研究 hESC 命运决定中涉及的分子机制的非常有用的工具。对于这些类型的研究,hESC 的饲养细胞依赖性培养通常是有问题的,因为饲养细胞的生理学受到药物处理或基因修饰的干扰,这可能会使研究结果变得模糊。在这项研究中,我们评估了三种常用的无饲养细胞培养条件,以确定它们是否支持 hESC 的未分化生长,并确定在这些条件下生长的 hESC 是否显示出与依赖饲养细胞的 hESC 相似的基因表达模式。我们的结果表明,在三种无饲养细胞条件下生长的 hESC 表达未分化标记基因的强度与在依赖饲养细胞的培养物中生长的 hESC 相同。此外,全基因组基因表达谱表明,在无饲养细胞或依赖饲养细胞培养条件下生长的 hESC 的基因表达模式高度相似。这些结果表明,无饲养细胞培养条件能够像依赖饲养细胞的培养条件一样有效地支持 hESC 的未分化生长。因此,无饲养细胞培养条件可能适合药物筛选和基础研究中 hESC 的基因操作。