人胎肺成纤维细胞对小鼠胚胎干细胞生长支持作用的研究

Human embryonic lung fibroblasts as feeder cells supporting the growth of mouse embryonic stem cell

目的探讨人胎肺成纤维细胞(hELF)作为饲养层支持小鼠胚胎干细胞(ES)的生长及维持ES细胞的未分化状态。方法采用生长状态良好的hELF,经一定浓度的丝裂霉素C处理后,制备hELF饲养层,将ES细胞接种到该饲养层进行传代培养。观察ES细胞的形态学特征,测定ES细胞表面碱性磷酸酶(AKP)活性、干细胞标志物Oct-4的表达。结果 hELF细胞经丝裂霉素C处理后不再增殖,保持较好的细胞功能和形态学特征。ES细胞克隆在饲养层呈现集落状隆起生长,边缘清楚,结构致密,与周围的饲养层细胞界限清楚。ES细胞仍然保持未分化状态,高度表达AKP及胚胎干细胞转录因子Oct-4。结论 hELF作为饲养层可以较好的维持胚胎干细胞的生长及其未分化状态。hELF源自人工流产组织,取材来源较丰富,为hELF饲养层应用于人胚胎干细胞的培养奠定了基础。

Objective： To explore whether human embryonic lung fibroblasts （hELF）could be used as feeder layer to support the growth of embryonic stem cells （ESC） and keep ESC undifferentiated. Methods： hELFs were treated by optional concentrations of mitomycin-C and prepared as feeder layer on which ESCs were transplanted for subculturing. The morphological characteristics of ESCs, alkaline phosphatase （AKP） activity and the expression of stem cell marker Oct-4 in ESCs were observed. Results.. The proliferation of hELFs can be inhibited by mitomycin-C, but hELFs can maintain good function and morphological characteristics. The ESC clones looked like islands on the feeder, with clear and smooth edge. There was no clear boundary between ESCs. The ESCs remained undifferentiated, with high expression of AKP and Oct 4. Conclusions： hELFs which were obtained from aborted fetal tissues could serve as a convenient resource of feeder layer to effectively support the growth of ESCs.