Feeder cells provide an optimal microenvironment for the propagation of human embryonic stem cells (hESCs) by supplying currently known or unknown factors. However, the hESCs grown on feeder cells are not suitable for the purpose of clinical application because of the risk of contamination. In recent years, the feeder-free culture method has been developed to eliminate contamination, but some studies show that hESCs exhibit poor growth patterns in a feeder-free culture system. Regarding this phenomenon, we speculate that some genes related to hESC propagation were differently expressed in hESCs grown on feeder cells. To test this hypothesis, 3 hESC lines (NF4, NF5 and P096) were efficiently expanded in a feeder-free culture system or on human foreskin fibroblast (HFF) cells. The different gene expression patterns of hESCs in these 2 conditions were analyzed through microarrays. The results revealed that the hESCs cultured in both conditions maintained the expression of stemness markers and the ability to spontaneously differentiate into the 3 germ layers. The analysis of gene expression profiles revealed that 23 lncRNA and 15 genes were significantly differentially expressed in these two culture conditions. Furthermore, GO analyses showed that these genes were involved in such biological processes as growth factor stimuli, cell growth, and stem cell maintenance. To summarize, our study demonstrated that the hESCs grown on the HFF showed different gene expression patterns compared to those grown in a feeder-free culture system, suggesting that these differently expressed lncRNAs and genes played important roles in maintaining hESC propagation.

中文翻译：

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在无饲养层培养系统和人包皮成纤维细胞饲养层中生长的人类胚胎干细胞的不同转录谱

饲养细胞通过提供目前已知或未知的因子，为人类胚胎干细胞 (hESC) 的繁殖提供最佳微环境。然而，由于存在污染风险，在饲养细胞上生长的 hESC 不适用于临床应用。近年来，开发了无饲养层培养方法来消除污染，但一些研究表明，hESCs 在无饲养层培养系统中表现出较差的生长模式。关于这种现象，我们推测一些与 hESC 繁殖相关的基因在饲养细胞上生长的 hESC 中表达不同。为了检验这一假设，3 个 hESC 细胞系（NF4、NF5 和 P096）在无饲养层培养系统或人包皮成纤维细胞 (HFF) 中有效扩增。通过微阵列分析了这两种情况下 hESC 的不同基因表达模式。结果表明，在两种条件下培养的 hESC 均保持了干性标记物的表达和自发分化为 3 个胚层的能力。基因表达谱分析显示，23 个 lncRNA 和 15 个基因在这两种培养条件下显着差异表达。此外，GO 分析表明，这些基因参与了诸如生长因子刺激、细胞生长和干细胞维持等生物过程。总而言之，我们的研究表明，与在无饲养层培养系统中生长的 hESC 相比，在 HFF 上生长的 hESC 表现出不同的基因表达模式，