肝癌患者脂肪干细胞重编程为诱导多潜能干细胞  

Reprogramming of adipose-derived stem cells of hepatic cancer patients into pluripotent stem cells

作　　者：付玉华[1] 范丽[1] 丁丽[1] 陈艳[2] 周秀梅[1] 王欢[1] 刘韬[2] 王敬晗[2] 李琳芳[2] 钱其军[1,2] 

机构地区：[1]浙江理工大学生命科学学院,新元医学与生物技术研究所,杭州310018 [2]第二军医大学东方肝胆外科医院基因-病毒治疗实验室,上海200438

出　　处：《第二军医大学学报》2011年第4期382-386,共5页Academic Journal of Second Military Medical University

基　　金：国家杰出青年科学基金(30925037)~~

摘　　要：目的重编程肝癌患者来源的脂肪干细胞为诱导多潜能干细胞。方法包装携带Oct4、Sox2、Klf4、c-Myc基因的反转录病毒,将病毒感染脂肪干细胞并培养诱导后的细胞,采用碱性磷酸酶染色、定量PCR和原位免疫荧光实验鉴定诱导的克隆样细胞。结果诱导的克隆样细胞表达碱性磷酸酶,定量PCR证实克隆样细胞表达胚胎干细胞多能性基因,免疫荧光实验证实其表达Oct4和Sox2。结论肝癌患者来源的脂肪干细胞可高效重编程为诱导多潜能干细胞,且脂肪干细胞可作为培养诱导多潜能干细胞的滋养细胞,为提高成体细胞重编程效率和建立基于诱导多潜能干细胞的肝癌模型研究提供了平台。Objective To reprogram the adipose-derived stem cells of hepatic cancer patients into induced pluripotent stem cells（iPS cells）.Methods Retrovirus-based pMXs containing Oct4,Sox2,Klf4 and c-Myc were prepared and were used to infect adipose-derived stem cells;the infected cells were then cultured.Alkaline phosphatase staining,quantitative-PCR and immunofluorescence assay were used to identify the ES-cell-like colonies. Results The ES-cell-like colonies expressed Alkaline phosphatase.Quantitative-PCR showed that ES-cell-like cells expressed the pluripotency genes of ES cells.Furthermore,immunofluorescence confirmed the expression of Oct4 and Sox2 in ES-cell-like cells.Conclusion Adipose-derived stem cells of hepatic cancer patients can be efficiently reprogrammed into iPS cells,and they can serve as the feeder layers of their own iPS cells.These findings may pave a way for improving efficiency of reprogramming adult cells and establishing hepatic cancer model for inducing pluripotent stem cells.

关 键 词：肝肿瘤 脂肪干细胞 诱导多潜能干细胞 重编程 反转录病毒 

分 类 号：R329.29[医药卫生—人体解剖和组织胚胎学]