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目的观察胚胎干细胞(ESC)来源的肝细胞对急性肝功能衰竭(FHF)小鼠的移植治疗方法和作用。方法选用小鼠D3-ES细胞,利用转化生长因子、成纤维细胞生长因子、肝细胞生长因子等进行肝细胞方向诱导分化;用反转录聚合酶链反应、免疫细胞化学等方法检测肝细胞标记物甲胎蛋白(AFP)、白蛋白(ALB)和葡萄糖6磷酸酶(G6P)等的表达;观察阳性细胞的空间分布规律,用细胞铲分选阳性区域细胞并将其移植入同系(129系)小鼠肝脏内,利用四氯化碳诱发FHF,然后观察小鼠生存时间、肝功能指标和移植细胞生长状态等情况。结果(1)AFP、白蛋白和G6P等最早于第3天开始表达并逐渐增强。(2)移植组平均生存时间(60h)与对照组(22h)差异有统计学意义(P<0·05);移植组肝功能指标均较对照组明显改善(P<0·05)。(3)移植细胞表达ALB,移植部位未发现肿瘤形成。结论ESC来源的肝细胞可表达成熟肝细胞功能,移植后能对小鼠FHF起到较好的治疗作用。
Objective To observe the methods and effects of embryonic stem cell (ESC) -induced hepatocytes transplantation in acute liver failure (FHF) mice. Methods Mouse D3-ES cells were selected and induced to differentiate into hepatocytes using transforming growth factor, fibroblast growth factor and hepatocyte growth factor. Reverse transcription-polymerase chain reaction and immunocytochemistry were used to detect hepatocyte markers (AFP), albumin (ALB) and glucose 6-phosphatase (G6P) were detected by immunohistochemistry. The spatial distribution of positive cells was observed. Positive cells were screened by the cell shovel and transplanted into homologous line 129 ) Mouse liver, carbon tetrachloride-induced FHF, and then observed the survival time of mice, liver function indicators and cell growth status. Results (1) AFP, albumin and G6P, etc. expressed as early as the third day and gradually increased. (2) The mean survival time of the transplantation group (60h) was significantly different from that of the control group (22h) (P <0.05). The liver function of the transplantation group was significantly improved compared with the control group (P <0.05). (3) The transplanted cells expressed ALB, and no tumor formation was found in the transplanted area. Conclusions ESC-derived hepatocytes express mature hepatocyte function, and have good therapeutic effect on mouse FHF after transplantation.