免疫磁珠分选兔前软骨干细胞及细胞研究平台的建立  

Immunomagnetic bead separation and culturing of precartilagious stem cells from fetal rabbit

作　　者：张瑞[1] 王俊芳[1] 程力[1] 

机构地区：[1]南京医科大学附属无锡市人民医院骨科,江苏无锡214023

出　　处：《中国矫形外科杂志》2013年第13期1352-1358,1356-1358,共7页Orthopedic Journal of China

基　　金：南京医科大学科技发展基金重点项目(项目编号:2010NJMUZ64;No.2011NJMU205);国家自然科学基金青年科学基金项目(项目编号:81101374)

摘　　要：[目的]研究兔前软骨干细胞PSCs,(precartilagious stem cells)的分离、培养方法及增殖、表型特征,为细胞移植治疗椎间盘退变,提供合适的种子细胞。[方法]取胎兔骨骺周围软骨膜(LaCroix环)中的细胞进行原代培养,然后采用免疫磁珠分离系统分选纯化PSCs。体外培养,传代,冻存复苏,绘制生长曲线,观察细胞特性。分别采用免疫组化、免疫荧光、RT-PCR等方法鉴定纯化后的PSCs。[结果]免疫磁珠分离可获得纯度较高的兔PSCs,培养后成活率高,细胞状态良好。细胞冻存复苏后,细胞增殖速度、细胞形态及表面标志物无明显变化。免疫组化、免疫荧光、RT-PCR等方法鉴定后,细胞都有明显的PSCs表面特异性标记物的表达。[结论]PSCs存在于LaCroix环中,免疫磁珠分离法得到的PSCs,体外培养条件下,细胞增殖较快,生物学特性稳定。[ Objective ] To isolate and culture rabbit precartilagious stem cells （PSCs） and to investigate their phenotypic characteristics and cell transplantation for further related research. ~ Methods] PSCs were separated and purified from the peri- ehondrium around the epiphyseal of fetal rabbits, cultured in vitro, passaged, and then eryopreserved. We plotted the growth curve for the cultured PSCs and observed their characteristics. Immunohistochemistry, immunofluorescence, and RT- PCR ana- lyses were used to identify purified PSCs. [ Results] Using the immunomagnetic separation method, we obtained high -purity rabbit PSCs showing high survival rate after culture, and these purified cells were in good condition. The cryopreserved cells did not show significant changes in the rate of proliferation, morphology, and surface markers profiles. Immunohistochemistry, immu- nofluorescenee, and RT - PCR analyses revealed that the purified cells expressed PSC - specific surface markers. [ Conclu- sion] PSCs exist in the LaCroix ring, The PSCs isolated using the immunomagnetic separation method showed rapid cell prolifer- ation and stable biological characteristics.

关 键 词：前软骨干细胞 免疫磁珠分选 成纤维细胞生长因子受体-3 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]