骨形态发生蛋白9-2双表达定向诱导多潜能干细胞C3H10的成骨分化  

Co-expression of bone morphogenetic protein 9-2 induces differentiation of multipotential stem cells C3H10 into osteoblasts

作　　者：陈聪[1] 罗庆[1] 田雯[1] 迭小红[1] 康权[1] 

机构地区：[1]重庆医科大学附属儿童医院 儿童发育疾病研究省部共建教育部重点实验室 儿科学重庆市重点实验室 重庆市儿童(发育重大疾病诊治与预防国际科技合作基地,重庆400014

出　　处：《中国生物制品学杂志》2013年第1期32-35,共4页Chinese Journal of Biologicals

基　　金：国家自然科学基金(30973062);重庆市自然科学基金(CSTC2008BB5390;CSTC2009BB5408)

摘　　要：目的探讨骨形态发生蛋白(Bone morphogenetic protein,BMP)9-2双表达定向诱导多潜能干细胞C3H10向成骨细胞分化的情况。方法将C3H10细胞分为4组:BMP9-2、BMP2、BMP9和GFP组,将4种重组腺病毒分别感染C3H10细胞,通过碱性磷酸酶(Alkaline phosphatase,ALP)染色、定量测定及钙茜素红染色,观察BMP9-2双表达对C3H10细胞成骨分化的定向诱导作用。结果 BMP9-2双表达能诱导C3H10细胞ALP的表达,其染色活性高于BMP2组1.6倍,BMP9组2.5倍,GFP组4倍;其BMP9-2双表达定量表达A520值在病毒感染后5、7、9 d均高于BMP2、BMP9和GFP组;BMP9-2双表达能诱导C3H10细胞钙盐的沉积,感染14 d后,钙茜素红染色可见钙化结节,其染色活性高于BMP2组1.4倍,BMP9组1.3倍,GFP组5.2倍。结论 BMP9-2双表达能定向诱导C3H10细胞成骨分化,其成骨活性强于单一成骨诱导因子BMP2和BMP9。Objective To investigate the capacity of co-expression of bone morphogenetic protein 9-2（BMP9-2） in inducing the differentiation of multipotential stem cells C3H10 into osteoblasts.Methods C3H10 cells were divided into four groups,and infected with recombinant adenoviruses with BMP9-2,BMP2,BMP9 and GFP respectively.The induction of differentiation of C3H10 cells to osteogenesis by co-expression of BMP9-2 was observed by alkaline phosphatase（ALP） staining,alkaline phosphatase quantitation and calcium-alizarin red staining.Results The co-expression of BMP9-2 induced the expression of ALP in C3H10 cells.The positive rate of ALP in BMP9-2 group was 1.6 times higher than that in BMP2 group,2.5 times higher than that in BMP9 group,and 4 times higher than that in GFP group.The activities（A520） of ALP in BMP9-2 group 5,7 and 9 d after infection were higher than those in BMP2,BMP9 and GFP groups.The co-expression of BMP9-2 induced the sedimentation of calcium salt in C3H10 cells.Calcified tuberculum was observed in C3H10 cells 14 d after infection with BMP9-2 by calcium-alizarin red staining.The sedimentation level in BMP9-2 group was 1.4 times higher than that in BMP2 group,1.3 times higher than that in BMP2 group,and 5.2 times higher than that in GFP group.Conclusion The co-expression of BMP9-2 may induce the differentiation of C3H10 cells into osteoblasts,of which the ability in inducing osteogenesis is higher than those of BMP2 and BMP9.

关 键 词：骨形态发生蛋白质类 双表达 多潜能干细胞 C3H10细胞 成骨 重组腺病毒 

分 类 号：Q254[生物学—细胞生物学]