DA-2006促进骨髓基质干细胞分化为成骨细胞效果及机制分析

摘要

Objective:In vitro to study on rabbit bone marrow stromal stem cells into osteoblasts differentiation effects and molecular mechanism. Method:We obtained rabbit femur, and flush the bone marrow cavity. We adopt the whole bone marrow culture method for the separation and puriifcation of BMSCs. When BMSCs grows as the third generations, we respectively use the induction culture medium(Group A), the common culture medium contained DA-2006(Group B), and the common culture medium(the control group). MTT were used in three groups of bone marrow stromal stem cell proliferative capacity. We adopt the lfuorescence quantitative PCR technique for the detection of bone alkaline phosphatase (ALP), osteocalcin (OC), osteopontin gene expression as osteogenic markers. Results:BMSCs grows as better morphologyand quickly. In the growth rate, the group B had strong that was better than the group A, the lowest blocking level was the control group. the ALP, OC, OPN expression of the Group B were signiifcantly higher than group A and the control group(P<0.05). Conclusion:In the containing DA-2006 cell culture medium induced environment, the promotes the osteoblast differentiation from rabbit bone marrow stromal stem cells is better than the classic induced by dexamethasone, and its mechanism may play up ALP, OC and OPN expression levels.%目的：探讨体外培养条件下DA-2006促进骨髓基质干细胞分化为成骨细胞的效果与机制。方法：取兔股骨,骨髓腔进行冲洗,采用全骨髓法分离纯化培养BMSCs,培养至第3代后，分别利用诱导培养液(组A )，添加D A-2006的培养液(组B )以及普通培养液(对照组)培养BMSCs，采用噻唑蓝(MTT)检测3组骨髓基质干细胞增殖能力,利用荧光定量PCR技术检测骨碱性磷酸酶(ALP)、骨钙素(OC)、骨桥蛋白(OPN)等成骨性标志基因的表达情况。结果：经过诱导后，组B细胞具有良好的形态，扩增能力迅速。在生长率方面，诱导组B较强于诱导组A，对照组团块化程度最低。对照组的表达量最低，同时组B的ALP、OC、OPN表达量明显高于组A ( P<0.05)。结论：在含D A-2006的细胞培养液诱导环境下，骨髓基质干细胞的成骨分化强于经典的地塞米松诱导效果，其机制的发挥可能与上调ALP、OC与OPN的表达水平有关。