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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 adipose-derived stem cells from patients with hepatocellular carcinoma to induce pluripotent stem cells. Methods Retroviruses carrying Oct4, Sox2, Klf4 and c-Myc genes were packaged and infected with ADSCs. The induced cells were cultured. The induced clones were identified by alkaline phosphatase staining, quantitative PCR and in situ immunofluorescence assay Like cells. Results The induced cloned cells expressed alkaline phosphatase. Quantitative PCR confirmed that clonal cells expressed pluripotency genes of embryonic stem cells. The expression of Oct4 and Sox2 was confirmed by immunofluorescence. Conclusion ADSCs derived from hepatocellular carcinoma patients can be efficiently reprogrammed to induce pluripotent stem cells. Adipose-derived stem cells can be used as trophoblasts to induce induced pluripotent stem cells. To provide the basis for improving the reprogramming efficiency of adult cells and establishing a hepatoma model based on induced pluripotent stem cells The platform.