小鼠胚胎成纤维细胞重编程为多能干细胞

背景：目前胚胎干细胞研究面临伦理争议及来源困难等诸多问题,人们一直在寻找不需破坏胚胎或卵细胞的建立多潜能干细胞的方法。 目的：构建小鼠诱导性多潜能干细胞系并对其进行鉴定。 方法：利用反转录病毒将3个干细胞基因Oct4、Sox2、Klf4导入小鼠胚胎成纤维细胞中,将其诱导为多潜能干细胞。通过镜下观察、碱性磷酸酶染色、反转录PCR(RT-PCR)、免疫荧光实验及畸胎瘤形成实验等对诱导性多潜能干细胞形态、多能性基因表达情况、干细胞表面标记及全能性等进行鉴定分析。 结果与结论：从小鼠胚胎成纤维细胞诱导而来的诱导性多潜能干细胞镜下观察呈典型的克隆状生长,圆形或椭圆形,与饲细胞分界清楚;RT-PCR、碱性磷酸酶染色及免疫荧光检测诱导性多潜能干细胞高表达胚胎干细胞相关基因及蛋白;种植在免疫缺陷鼠体内能够形成向内中外3个胚层分化的畸胎瘤,表明诱导性多潜能干细胞具有多潜能性。通过转导3种重编程因子可以将小鼠成纤维细胞诱导为类似胚胎干细胞的多潜能干细胞。

Induction of pluripotent stem cells from mouse embryonic fibroblast cultures by three defined factors

BACKGROUND:The study of embryonic stem cells is always confined to ethical disputation or lack of ootid. Scholars are trying to seek a method to obtain multi-potential stem cells which would not destroy the embryo or egg cells. OBJECTIVE:To construct and identify the induced pluripotent stem cells from mouse embryonic fibroblasts by three defined factors. METHODS:Three stem cell genes (Oct4, Sox2 and Klf4) were introduced into the embryonic fibroblasts of BALB/C mouse by retroviral infection. The induced pluripotent stem cells constructed by this method were analyzed in many aspects, including surface antigens, gene expression, alkaline phosphatase activity and teratoma formation assays． RESULTS AND CONCLUSION:The induced pluripotent stem cells generated from this study had an embryonic stem cell-like shape, were positive for alkaline phosphatase staining, expressed embryonic stem cell-specific surface antigens, and possessed the ability to differentiate into 3-germ layers both in vivo and in vitro. So the induced pluripotent stem cells can be obtained from mouse embryonic fibroblasts by introducing three factors, which are similar to embryonic stem cells in many ways.