Objective: To investigate the effect of basic fibroblast growth factor (bFGF) secreted by human or mouse feeder fibroblasts on the undifferentiated growth of human embryonic stem cells (hESCs).rnMethods: The hESCs were cultured with feeder cells which consisted of human or mouse fibroblasts. The Enzyme-linked immunosorbent assay was applied to analyze the expression levels of bFGF by human and mouse feeder cells.rnResults: The adherent efficiency and clonal growth rate were not significantly different between human and mouse feeder cells (P>0. 05). The undifferentiated rate of the third generation of hESCs supported by the mouse feeder cells was higher than that of by human feeder cells(P<0. 05). It was found that human feeder cells secreted larger amount of bFGF, but bFGF in culture media from mouse feeder cells could not be detected.rnConclusions: The fibroblast as feeder cells could promote the undifferentiated growth of hESCs. The bFGF secreted by human fibroblasts increased the adherence and growth of hESCs.%目的 初步探讨人源性和鼠源性饲养层细胞分泌的碱性成纤维生长因子(bFGF)对人胚胎干细胞(hESCs)未分化生长的影响. 方法 人源性和鼠源性成纤维细胞制备成饲养层(人源性饲养层不加bFGF,鼠源性饲养层加4ng/ml bFGF),同期接种同一株hESCs,对比hESCs的贴壁率、克隆生长率及未分化率,比较它们支持hESCs生长的差异;通过酶联免疫吸附实验(ELISA)方法,快速检测评估人源性和鼠源性饲养层细胞自身分泌的bFGF. 结果 人包皮和鼠源性饲养层培养hESCs的贴壁率和克隆生长率之间无显著差异(P＞0.05),但是鼠源性支持三代hESCs的未分化率均高于人源性饲养层(P＜0.05).鼠源性饲养层不能分泌任何可测量到的bFGF,而人源性饲养层细胞均能分泌bFGF. 结论 饲养层细胞自身分泌的bFGF能支持hESCs的贴壁和增殖生长,但不同饲养层支持hESCs生长的最适宜bFGF浓度和抑制hESCs分化的作用机制,还有待于进一步研究.
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