骨形态发生蛋白-2、-4和-6在脂肪间充质干细胞向软骨细胞诱导中的调节作用  

Regulating effect of bone morphogenetic protein-2, -4, and -6 on chondrocyte formation from adipose derived stem cells

作　　者：但洋[1] 刘鹏[2] 刘琦斌[1] 余慧娟[1] 郑东[1] 杨述华[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院骨科,武汉430022 [2]华中科技大学同济医学院附属协和医院胸外科,武汉430022

出　　处：《中华实验外科杂志》2014年第4期808-811,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81101376）

摘　　要：目的观察骨形态发生蛋白（BMP）-2、BMP-4和BMP-6对兔脂肪间充质于细胞（ADSCs）向软骨细胞分化过程中的影响。方法流式细胞仪鉴定脂肪干细胞表面抗原CD34、CD45、CD44和CD29，实验组分别增加100ng/L的BMP-2、BMP-4和BMP-6，与转化生长因子（TGF）-β3-起在软骨细胞诱导液中进行诱导。21d后，阿利新蓝染色检测酸性糖胺聚糖，实时定量聚合酶链反应（Real-timePCR）和Westernblot检测聚集蛋白聚糖（ACAN）、II型胶原（COL2A1）和性别决定区Y框蛋白-9（SOX-9）的mRNA和蛋白的表达。结果BMP-4组仅增加了细胞增殖，BMP-6组比BMP-2组，明显提高细胞蛋白多糖的表达，BMP-6组比BMP-2组ACAN、COL2A1和SOX-9mRNA表达分别增加1．7、1．3和1．0倍，蛋白表达分别增加1．4、1．3和0．9倍。结论TGF-β3与BMP-6组合更能增加ADSCs向软骨细胞分化的效率。Objective To observe the influence of cytokines of bone morphogenetic protein （BMP）-2, -4, and -6 on the process of ehondrocyte formation from rabbit adipose derived stem cells （ADSCs）. Methods The surface antigens CD34, CD45, CIM4 and CD29 of ADSCs were identified by flow cytometry, and ADSCs were cultured for 21 days by using the chondrocyte inducing conditional media serving as the cuhure media containing transforming growth factor （TGF）-β3, and respectively 100 μg/L BMP-2, BMP-4 or BMP-6 to experiment groups. Acidic glycosaminoglycans were detected by alcian blue staining. The mRNA and protein expression levels of aggreean （ACAN）, type II collagen （COL2A1） and sex determining region Y-box 9 （SOX-9） were analyzed by real-time quantitative polymerase chain reaction （Real-time PCR）, and Western blotting, respectively. Results Only cell proliferation is realized in BMP-4 group, while cell proteoglycan expression levels were more obviously raised in BMP-6 group than in BMP-2 group. The mRNA expression levels of ACAN, COL2A1, and SOX-9 in BMP-6 group were separately 1.7, 1.3 and 1.0 times higher than those in BMP-2 group. And protein expression levels of ACAN, COL2A1, and SOX-9 in BMP-6 group were 1.4, 1.3 and 0. 9 times higher than those in BMP-2 group. Conclusion The combination of TGF-β3 and BMP-6 can promote higher efficiency of differentiation of ADSCs into chon- drocytes than BMP-2 and BMP-4.

关 键 词：脂肪干细胞 骨形态发生蛋白 软骨细胞 转化生长因子-3 组织工程 

分 类 号：R346[医药卫生—基础医学]