骨骺干细胞复合多孔β-磷酸三钙支架体外培养的实验研究  

Experimental studies on precartilaginous stem cells cultured withβ-tricalcium phosphate scaffold in vitro

作　　者：谢虎 郭风劲[2] 张继平 李永忠 孙淑珍[2] 祁军[3] 

机构地区：[1]湖北省宜昌市第二人民医院骨科,宜昌443000 [2]华中科技大学附属同济医院骨科 [3]武汉理工大学材料学院

出　　处：《中华小儿外科杂志》2012年第2期136-138,共3页Chinese Journal of Pediatric Surgery

基　　金：国家自然科学基金（批准号：30571872）

摘　　要：目的 观察免疫磁珠分选的骨骺干细胞（PSCs）诱导后向软骨特性方向分化并与多孔β-磷酸三钙（β-TCP）材料复合体外培养,探讨其构建组织工程化骺板的可行性.方法 免疫磁珠分选系统分离纯化PSCs体外培养并用免疫细胞化学方法对其进行鉴定,诱导其向软骨细胞特性方向分化,免疫荧光染色及甲苯胺兰染色检测诱导后细胞Ⅱ型胶原及软骨特征基质表达情况.诱导后的PSCs与多孔β-磷酸三钙支架复合培养后扫描电镜观察其粘附增殖及形态学改变.结果 免疫磁珠系统分选的PSCs体外培养增殖迅速,阳性细胞率超过90％,诱导后的PSCsⅡ型胶原免疫荧光染色及甲苯胺兰染色阳性.细胞接种支架后在多孔β-磷酸三钙支架表面生长增殖良好并分泌细胞外基质.结论 免疫磁珠分选系统分离的PSCs经诱导后向软骨细胞功能方向分化,与多孔β-磷酸三钙支架复合有望构建组织工程化骺板.Objective To observe the biological behavior of cultured pre-cartilaginous stem cells （PSCs） combined with β-tricalcium phosphate（β-TCP） scaffold and to investigate its feasibility for epiphyseal plate tissue engineering.Methods PSCs were isolated and purified using immunomagnetic microbeads,and were induced into chondrogenic differentiation.Collagen Ⅱ were detected by immunofluorescence staining,and extracellular matrix was detected by Toluidine Blue staining.Then the cells were seeded onto the scaffold and the effects of adhesion and morphological changes were observed by the phase-contrast microscopy and scanning electron microscopy （SEM）.Results The PSCs were successfully cultured in vitro.The induced cells showed positivity for Collagen Ⅱ and positive staining of extracellar matrix,and adhered to the surface of the scaffold and proliferated well.Conclusions PSCs could be effectively isolated and purified by immunomagnetic microbeads and induced into chondrogenic differentiation,and β-TCP scaffold loaded PSCs may be used in epiphyseal plate tissue engineering.

关 键 词：骨骺干细胞 Β-磷酸三钙 骺板 组织工程 

分 类 号：R726.8[医药卫生—儿科]