【摘要】 目的建立无血清无饲养层人胚胎干细胞(hESCs)胶质前体细胞分化体系。方法将生长在层黏连蛋白包被的培养板上的人胚胎干细胞,悬浮培养诱导拟胚体(EBs)形成,将25d的EBs转移至含有胰岛素、5μg/L碱性成纤维细胞生长因子(bFGF)、20μg/L表皮生长因子(EGF)和5μg/L三碘甲状腺素(T3)的胶质细胞分化培养基贴壁培养7～10d,诱导胶质前体细胞分化。收集成纤维样分化细胞,采用RT-PCR法检测胶质前体细胞早期基因的表达,细胞免疫荧光染色检测胶质前体细胞标记的表达。结果分化得到的细胞呈长梭形,表达神经胶质前体细胞标记NG2和血小板衍生生长因子受体α(PDGFRα),阳性率分别为37.2%和47.6%。该前体细胞可进一步分化为星形胶质细胞和少突胶质细胞,未见神经元。结论含有胰岛素、bFGF、EGF和T3的胶质细胞分化培养基可诱导hESCs向胶质前体细胞分化。更多还原

【Abstract】 Objective To establish a neuroglia progenitor differentiation system from human embryonic stem cells(hESCs) in a feeder layer-and serum-free medium.Methods hESCs grown in laminin coated culture plates were induced to form embryoid bodies(EBs) for 25 days.Then 25-day-old EBs were transferred to glial progenitor induction medium(GPIM) containing insulin,5μg/L basic fibroblast growth factor(bFGF),20μg/L epidermal growth factor(EGF) and 5μg/L triiodothyronine(T3) to continue culturing for 7-10 days.The differentiated fibroblast-like cells were digested and collected.The expressions of neuroglia progenitor genes were determined by RT-PCR.The special markers of neuroglia progenitors were examined by immunofluorescence staining.Results The differentiated cells were long spindle shaped.They expressed neuroglia progenitor special markers NG2,platelet-derived growth factor receptor α(PDGFRα).The percentage of NG2 +,PDGFRα + cells was 37.2% and 47.6%,respectively.They could further differentiate into astrocytes and oligodendroglias.Conclusion GPIM containing insulin,bFGF,EGF and T3 could induce hESCs differentiate into neuroglia progenitors.This in vitro system may be useful for studying mechanism of neuroglia differentiation.更多还原