肝损伤大鼠血清诱导培养脐带间充质干细胞向肝样细胞的分化

Serum from liver injury rats induces differentiation of umbilical cord mesenchymal stem cells into hepatocyte-like cells

背景:脐带间充质干细胞来源于新生个体脐带组织,来源广泛,无社会伦理学因素制约,有望代替骨髓间充质干细胞成为细胞移植和再生医学的理想种子细胞。目的:探讨肝衰竭大鼠血清对人脐带间充质干细胞肝向诱导分化作用,为临床上利用脐带间充质干细胞治疗终末期肝病提供实验依据。方法:腹腔注射10%四氯化碳溶液建立急性肝损伤大鼠模型,对照组腹腔注射等剂量的大豆油,48 h后腹主动脉取血离心分离血清。取第3代人脐带间充质干细胞,分别用体积分数为20%肝损伤大鼠血清和体积分数为20%胎牛血清诱导培养,观察诱导前后人脐带间充质干细胞形态学改变,检测培养液上清甲胎蛋白、白蛋白水平。结果与结论:肝损伤大鼠血清培养第1天细胞形态变化不大,呈梭形,仍呈编织状或漩涡状生长,第2天细胞呈短梭形,第3天细胞呈类圆形,第4天呈圆形,并有极少量细胞漂浮。肝损伤大鼠血清培养人脐带间充质干细胞4 d,其上清液白蛋白水平较诱导前和对照组均有升高趋势(P<0.001),甲胎蛋白水平无明显变化。结果表明肝损伤大鼠血清可使脐带间充质干细胞向肝样细胞分化。

BACKGROUND： Umbilical cord mesenchymal stem cells are from the umbilical cord of newly born individuals and have no ethical issues, and therefore are promising candidates for seeded cells as a substitute for cell transplantation and regenerative medicine. OBJECTIVE： To investigate the effects of serum from liver injury rats on induced differentiation of human umbilical cord mesenchymal stem cells into hepatocyte-like cells and provide experimental evidence for use of human umbilical cord mesenchymal stem cells in the treatment of patients with end-stage liver disease in the clinic. METHODS： Rat models of acute liver injury were established by intraperitoneal injection of 10% carbon tetrachloride. Rats in the control group were intraperitoneally administered the same amount of soybean oil. Forty-eight hours after modeling, abdominal aorta blood was taken for serum preparation. Passage 3 human umbilical cord mesenchymal stem cells were cultured with 20% serum from liver injury rats and 20% fetal bovine serum. Morphology of human umbilical cord mesenchymal stem cells was observed before and after culture. Levels of α-fetoprotein and albumin in the supernatant were detected. RESULTS AND CONCLUSION： Cells exhibited shuttle-shaped appearance and grew in whirlpool-like manner at 1 day after culture with serum from liver injury rats, exhibited short shuttle-shaped appearance at 2 days, were oval-shaped at 3 days, and were round and an extremely small number of cells were floated at 4 days. At 4 days after culture with serum from liver injury rats, level of albumin in the cell supernatant was significantly increased than that before induction and that in the control group（P〈0.001）, and there was no significant difference in level of α-fetoprotein in the cell supernatant. These results suggest that serum of liver injury rats can induce differentiation of umbilical cord mesenchymal stem cells into hepatocyte-like cells.