Li Huazhuang, Zhou Yue, Guo Guoning
Third Military Medical University, Chongqing 400037, P. R. China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2006 Feb;20(2):121-4.
To observe effects of the core binding factor alpha1 (Cbfalpha1) in its promoting differentiation of the rabbit marrow mesenchymal stem cells (MSCs) into osteoblasts.
The rabbit marrow MSCs were isolated and cultured in vitro and were divided into 3 groups. In the control group, the marrow MSCs were cultured by DMEM; in the single inducement group, they were cultured by the condition medium (DMEM, 10% fetal bovine serum, dexamethasone 10 mmol/L, vitamin C 50 mg/L, and beta-GP 10 mmol/L); and in the experimental group, they were transfected with AdEasyl/Cbfalpha1, and then were cultured by the condition medium. The alkaline phosphatase (ALP) activity and the expression of osteocalcin as the osteoblast markers were measured with the chemohistological and immunohistochemical methods at 3 days, 1, 2, 3, and 4 weeks after inducement.
More than 90% MSCs were grown well in vitro. The GFP was positive in MSCs after their being transfected with AdEasyl/Cbfalpha1. The ALP activity and the expression of osteocalcin were significantly upregulated in the transfection group compared with those in the single inducement group and the control group at 1, 2, 3, and 4 weeks (P < 0.05). The mineralized node began to appear at 2 weeks in the experimental group and the single induction group, but did not appear in control group.
Cbfalpha1 can obviously promote differentiation of the rabbit marrow mesenchymal stem cells into the osteoblasts.
观察核心结合因子α1(Cbfalpha1)促进兔骨髓间充质干细胞(MSCs)向成骨细胞分化的作用。
分离兔骨髓MSCs并进行体外培养,分为3组。对照组采用DMEM培养骨髓MSCs;单诱导组采用条件培养基(DMEM、10%胎牛血清、地塞米松10 mmol/L、维生素C 50 mg/L和β-甘油磷酸钠10 mmol/L)培养;实验组转染AdEasyl/Cbfalpha1后,再用条件培养基培养。诱导后3天、1、2、3和4周,采用化学组织学和免疫组织化学方法检测碱性磷酸酶(ALP)活性及作为成骨细胞标志物的骨钙素表达。
90%以上的MSCs在体外生长良好。转染AdEasyl/Cbfalpha1后的MSCs中绿色荧光蛋白呈阳性。转染组在诱导后1、2、3和4周时,ALP活性及骨钙素表达均显著高于单诱导组和对照组(P < 0.05)。实验组和单诱导组在2周时开始出现矿化结节,对照组未出现。
Cbfalpha1能明显促进兔骨髓间充质干细胞向成骨细胞分化。