目的 观察红景天苷联合肝细胞生长因子(HGF)诱导脐血间充质干细胞(MSCs)向肝样细胞分化的作用.方法 将第5代脐血MSCs随机分成4组,A组不处理,B组分别加入5(B1)、10(B2)、20 μmol/L(B3)的红景天苷5 μL,C组分别加入5(C1)、10(C2)、20 ng/mL(C3)的HGF 5μL,D组分别加入B1+ C1(D1)、B2+ C2(D2)、B3+ C3(D3)的红景天苷和HGF.分别于培养第1、7、14、21天,光镜下观察细胞形态变化,并计数肝样细胞的数量；采用全自动生化仪检测细胞上清液中的谷丙转氨酶(ALT),溴甲酚绿法检测上清液中的白蛋白(ALB),观察肝样细胞的分泌功能.结果 A组细胞均呈长梭形,各时点无明显变化；B、C、D组第7天梭形细胞两端逐渐变短,第14、21天细胞由长梭形变为三角形、多角形或类圆形(肝样细胞).A组高倍镜下均未见肝样细胞,B、C、D组在第7、14、21天均可见肝样细胞(P均＜0.05),数量随时间延长而增加(P均＜0.05)；D组肝样细胞数量高于同时点的B、C组(P均＜0.05),并随药物浓度增加而增加；D2、D3与D1比较,P均＜0.05,而D3与D2比较,P＞0.05.A组各时点细胞上清液中均未检测到ALT、ALB,B、C、D组第14、21天均检测到ALT、ALB(P均＜0.05),并随时间延长而升高(P均＜0.05)；D组ALT、ALB水平均高于同时点的B、C组,且随药物浓度的增加而升高(P均＜0.05),D2、D3与D1比较,P均＜0.05,而D3与D2比较,P＞0.05.结论 与单用HGF、红景天苷相比,二者联合应用可增强脐血MSCs向肝样细胞分化的诱导作用,其中以10 μmol/L红景天苷联合10 ng/mL HGF的诱导效果最佳.%Objective To investigate the feasibility of the induction of mesenchymal stem cells derived from human umbilical blood into hepatocytes-like cells by salidroside and aepatocyte growth factor. Methods The human umbilical blood-derived mesenchymal stem cells were seperated into four groups and induced with different contents of salidroside and hepatocyte growth factor(HGF) in vitro. The cell morphology was observed under microscope at different times. Specific markers, such as alanine transaminase (ALT) and albumin (ALB) , were analyzed by immunocytochemistry method at the same time. Results The inducing differentiation rates of hepatocytes-like cell were improved greatly by salidroside and hepatocyte growth factor ( HGF) compared with group A ( all P < 0. 05 ) . The combination of 10 μmol/L salidroside and 10 ng/mL HGF was the most suitable for inducing differentiation of the human umbilical blood-derived mesenchymal stem cells among the experimental groups (all P <0.05). Conclusion Human umbilical blood-dederived mesenchymal stem cells are capable of differentitating into hepatocytes-like cells induced by salidroside and HGF in vitro.
展开▼
