骨形态发生蛋白对兔骨髓间充质干细胞成软骨分化的定向诱导作用

Cartilage differentiation of rabbit messenchymal stem cells induced by bone morphogenetic protein

目的研究骨形态发生蛋白(BMP)对兔骨髓间充质干细胞(MSCs)的定向诱导作用,探讨MSCs的成软骨能力。方法分离并培养兔MSCs,BMP诱导MSCs向软骨细胞分化,观察其形态学改变及碱性磷酸酶(ALP)活性。将诱导后的MSCs接种PGA无纺网体外培养2周,将MSCsPGA无纺网复合体植入裸鼠背部皮下,2个月后取材,进行组织学观察。结果经BMP诱导后,MSCs的形态由长梭形向多角形和三角形转化,群体倍增时间约为3.0d。诱导5、10d后,ALP活性为0.282±0.015,0.502±0.012,明显高于对照组0.265±0.010,0.315±0.021(P<0.01),ALP染色阳性。MSCsPGA无纺网复合体植入裸鼠后,组织学可见软骨陷窝,陷窝内可见核蓝染的成软骨细胞,证实MSCs具有体内成软骨能力。结论经BMP诱导的MSCs向软骨细胞分化和增殖,在裸鼠体内可形成软骨组织。

Objective To study the cartilage differentiation of rabbit messenchymal stem cells (MSCs) induced by bone morphogenetic protein (BMP).Methods The MSCs were separated and cultured from rabbit thighbone marrow.Growth kinetics was studied by growth curve.The MSCs had been cultured in the BMP medium for 10 days,and then the ALP activity was observed.The induced MSCs/PGA was implanted in the nude mice subcutaneously.The implants underwent the histological analysis after 2 months.Results The MSCs were successfully cultured and induced by the BMP.The growth curve showed that the time of population doubling was 3.0 days.ALP activity was 0.282± 0.015, 0.502± 0.012 in BMP group and 0.265± 0.010, 0.315± 0.021 in control group respectively after induction for 5 and 10 days (P< 0.01).The cells displayed fibroblast-like morphology.Histological evaluation showed the cartilage formation in 2 months in vivo.[WT5”HZ]Conclusion BMP can induce MSCs to differentiate chondral cells,and MSCs have the capability to generate cartilage.