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| 肝癌患者脂肪干细胞重编程为诱导多潜能干细胞 |
| 付玉华1,范丽1,于丽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12* |
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| (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) |
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| 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。 |
| 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 |
| DOI：10.3724/SP.J.1008.2011.0382 |
| 投稿时间：2011-01-13修订日期：2011-02-24 |
| 基金项目:国家杰出青年科学基金 (30925037). |
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| Reprogramming of adipose-derived stem cells of hepatic cancer patients into pluripotent stem cells |
| FU Yu-hua1,FAN Li1,YU Li1,CHEN Yan2,ZHOU Xiu-mei1,WANG Huan1,LIU Tao2,WANG Jing-han2,LI Lin-fang2,QIAN Qi-jun1 2﹡ |
| (1. Xinyuan Institute of Medicine and Biotechnology, College of Life Science, Zhejiang Sci-Tech University, Hangzhou 310018, Zhejiang, China;2. Laboratory of Viral and Gene Therapy, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai 200438, China) |
| Abstract: |
| ObjectiveTo reprogram the adipose-derived stem cells of hepatic cancer patients into induced pluripotent stem cells (iPS cells). MethodsRetrovirus-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. ResultsThe 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. ConclusionAdipose-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. |
| Key words: liver neoplasms adipose-derived stem cells induced pluripotent stem cells reprogramming retrovirus |
