联合表达BMP2和Sox9促进体外培养间充质干细胞成软骨分化  

Co-expression of BMP2 and Sox9 promotes chondrogenic differentiation of mesenchymal stem cells in vitro

作　　者：廖军义 周年[1] 林良波[1] 易世雄[1] 凡廷旭 赵辰[1] 胡宁[1] 梁熙[1] 司维柯[2] 黄伟[1] 

机构地区：[1]重庆医科大学附属第一医院骨科,重庆400016 [2]第三军医大学检验系临床血液教研室,重庆400038

出　　处：《南方医科大学学报》2014年第3期317-322,共6页Journal of Southern Medical University

基　　金：国家自然科学基金(81371972;31070875)~~

摘　　要：目的探讨联合表达骨形态发生蛋白2(BMP2)和Sox9对间充质干细胞(MSCs)成软骨分化的影响,为构建组织工程软骨提供实验依据。方法以小鼠胚胎骨髓MSCs(C3H10T1/2)为种子细胞,重组腺病毒BMP2诱导其分化,联用重组腺病毒Sox9过表达Sox9信号,重组腺病毒GFP作为对照。Real-Time PCR检测成软骨标志物Ⅱ型胶原(Col2a1)、蛋白聚糖(Aggrecan)、X型胶原(Col10a1)表达变化;Alcian blue染色了解细胞基质糖胺聚糖合成情况;免疫组化和Western blotting检测Col2a1合成情况。结果 BMP2能诱导C3H10T1/2细胞成软骨分化,当与Sox9联合作用时,Sox9可促进BMP2诱导的(Col2a1、Aggrecan、Col10a1)基因表达上调;促进细胞基质糖胺聚糖、Col2a1蛋白合成。结论 BMP2与Sox9联合作用可以促进体外培养MSCs成软骨分化,可能为组织工程软骨提供新的方法。Objective To investigate the effect of co-expression of bone morphogenetic protein 2 （BMP2） and Sox9 on chondrogenic differentiation of mesenchymal stem cells （MSCs） in vitro and provide experimental evidence for tissue engineering of cartilage. Methods Mouse embryonic bone marrow MSC C3H10T1/2 cells were infected with recombinant adenovirus expressing BMP2, Sox9 and green fluorescent protein （GFP） for 3-14 days, with cells infected with the adenovirus carrying GFP gene as the control. The mRNA expression of the markers of chondrogenic differentiation, including collagen type II （Col2a1）, aggrecan （ACAN）, and collagen type X （Col10a1）, were determined by real-time PCR. Alcian blue staining was used for quantitative analysis of sulfated glycosaminoglycan in the cellular matrix. The expression of Col2a1 protein was assayed by immunohistochemical staining and Western blot analysis. Results Adenovirus-mediated BMP2 expression induced chondrogenic differentiation of C3H10T1/2 cells. Overexpression of Sox9 effectively enhanced BMP2-induced expression of the chondrogenic markers Col2a1, aggrecan and Col10a1 mRNAs, and promoted the synthesis of sulfated glycosaminoglycan and Col2a1 protein in C3H10T1/2 cells. Conclusion Co-expression of BMP2 and Sox9 can promote chondrogenic differentiation of MSCs in vitro, which provides a new strategy for tissue engineering of cartilage.

关 键 词：骨形态发生蛋白2 SOX9 间充质干细胞 成软骨分化 BONE morphogenetic protein 2 SOX9 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]