人羊膜间充质细胞向成骨细胞诱导分化的实验研究  

Experimental study on the induced differentiation of human amnion mesenchymal cells into osteoblasts

作　　者：舒峻[1] 郭丽丽 张可华[1] 蔡哲[1] 程立明[3] 李荣旗 陈瀛[3] 潘琳[1] 欧华[2] 黄小杰[1] 张向利 徐扬[2] 高艳[2] 崔晓惠[1] 张红霞 刘佳新 

机构地区：[1]卫生部中日友好医院临床医学研究所,北京100029 [2]卫生部中日友好医院妇产科 [3]卫生部中日友好医院骨科 [4]北京市科润维德生物技术有限公司

出　　处：《中华整形外科杂志》2011年第5期362-367,共6页Chinese Journal of Plastic Surgery

基　　金：北京市海淀区科技项目（K20090023）

摘　　要：目的 观察人羊膜间充质细胞（humanamnionmesenchymalcells，hAMCs）体外诱导向成骨细胞分化，为骨组织工程提供种子细胞。方法从剖宫产后废弃的人羊膜组织分离培养hAMCs，经成骨细胞诱导条件培养基诱导后，对细胞形态特征、碱性磷酸酶、骨桥素、骨钙素表达以及I型胶原分泌进行观察和检测。结果原代培养的hAMCs形态呈长梭形或不规则形，呈均匀分布生长，传代后细胞体积略变大，约5～7d传代1次。经成骨细胞诱导培养15d后，hAMCs碱性磷酸酶、骨钙素、骨桥素的表达呈阳性，并且检测有I型胶原分泌。结论hAMCs易于体外分离培养及扩增，体外成骨细胞定向诱导的hAMCs具有典型的成骨细胞的形态和功能性特征，是良好的骨组织工程种子细胞。Objective To investigate the feasibility of inducing differentiation of the human amniotic mesenchymal cells （hAMCs） into osteoblasts in vitro, so as to provide the seed cells for bone tissue engineering. Methods The hAMCs were isolated from abandoned human amnion and cultured in osteogenie media to induce the osteogenic differentiation in vitro. After hAMCs were induced by osteogenie media for 15 days, morphological observation, immunocytochemistry and western blot were used to study the cellular morphology and expression of alkaline phosphatase（ALP） , type I collagen, osteopontin and osteoealein. Results The primary cultured hAMCs had long spindle shape or irregular shape, which were distributed evenly. The cells were usually subeultured in 5 or 7 days. After subculture, the cells became larger. After cultured by osteogenie media for 15 days, the hAMCs were detected to express ALP, osteoealcin and osteopontin, and secrete type I collagen. Conclusions The hAMCs are isolated, cultured and amplified easily in vitro. The induced differentiated ceils by osteogenic media have typical osteoblast morphological and functional characteristics, which can be used as seed cells for bone tissue engineering.

关 键 词：人羊膜间充质细胞 成骨细胞 组织工程 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]