摘要
背景:人脐带间充质干细胞具有多向分化潜能,经诱导后是否可向心肌细胞分化?目前的报道尚不多。目的:观察人脐带间充质干细胞经5-氮胞苷诱导分化后肌钙蛋白I的表达情况。方法:取第2代人脐带间充质干细胞标本,经消化、离心、沉淀后,分为7份,制成细胞悬液,调整细胞浓度为2×107L-1,分别加入浓度为80,40,20,10,5,2.5,0μmol/L5-氮胞苷于同样条件继续培养,作用24h后除去,继续培养4周。结果与结论:①人脐带间充质干细胞经5-氮胞苷处理24h之后,各组细胞都会出现部分死亡,失去典型的梭形形态,成为棍状或者柱状,40,80μmol/L组细胞形态变化明显。②免疫组化染色显示5,10,20,40,80μmol/L组诱导后2周心肌特异性肌钙蛋白I表达呈阳性,对照组与2.5μmol/L组心肌特异性肌钙蛋白I表达为阴性。选取5个高倍视野进行细胞计数,80,40,20,10μmol/L组间心肌样细胞的转化率差异无显著性意义,均明显高于5μmol/L组(P<0.05)。③超微结构观察显示经5-氮胞苷诱导后4周时部分人脐带间充质干细胞内可见典型的肌小节及心房颗粒。而未经5-氮胞苷诱导的对照组无以上改变。结果提示经5-氮胞苷诱导后人脐带间充质干细胞能够向心肌样细胞转化。
BACKGROUND： Human umbilical cord mesenchymal stem cells possess multi-differentiation potential. There are few studiesdescribing whether human umbilical cord mesenchymal stem cells can be induced to differentiate into cardiornyocytes. OBJECTIVE： To investigate cardiac troponin I expression in human umbilical cord mesenchymal stem cells after 5-azacytidine induction in vitro. METHODS： Passage 2 human umbilical cord mesenchymal stem cells were digested, centrifuged and deposited. Cell suspension was prepared and cell concentration was adjusted to be 2×10^7/L. Then cells were cultured for 24 hours after addition of various concentrations of 5-azacytidine （80, 40, 20, 10, 5, 2.5, 0 μmol/L）. After removal of 5-azacytidine, cells were cultured for another 4 weeks. RESULTS AND CONCLUSION： After treatment with 5-azacytidine for 24 hours, some cells died in each group, and typical fusiform appearance turned into stick-like or column-like appearance, especially in the 40 and 80 μmol/L 5-azacytidine groups. Immunohistochemical staining results showed that cardiac troponin I expression was detected in human umbilical cord mesenchymal stem cells in 5, 10, 20, 40, 80 μmol/L 5-azacytidine groups after induction for 2 weeks, but it was not detected in the control and 2.5 μmol/L 5-azacytidine groups. Five high-fold fields were selected for cell counting. The number of cardiomyocytes was not significantly different between 80, 40, 20, 10 μmol/L 5-azacytidine groups, but it was significantly higher compared to that in the 5 μmol/L 5-azacytidine group. After 5-azacytidine treatment for 4 weeks, typical sarcomeres and cardiac atrium particles were observed in some human umbilical cord mesenchymal stem cells. Such changes were not observed in the control group in which 5-azacytidine induction was not used. These findings suggest that human umbilical cord mesenchymal stem cells were induced to cardiomyocyte-like cells by 5-azacytidine.
出处
《中国组织工程研究》
CAS
CSCD
2013年第23期4212-4215,共4页
Chinese Journal of Tissue Engineering Research
关键词
干细胞
干细胞培养与分化
脐带间充质干细胞
5-氮胞苷
免疫组化
心肌细胞
肌钙蛋白I
干细
胞图片文章
stem cells
stem cell culture and differentiation
umbilical cord mesenchymal stem cells
5-azacytidine
immunohistochemistry
cardiomyocyte
cardiac troponin I
stem cell photographs-containing paper
