SD大鼠成骨细胞培养方法的改良  

Improvements of culture method of SD rat osteoblasts

作　　者：高蔚然[1] 袁亚江[1] 房师强[1] 梅晰凡[1] 

机构地区：[1]辽宁医学院附属一院骨科

出　　处：《解剖学杂志》2015年第2期162-164,194,共4页Chinese Journal of Anatomy

基　　金：辽宁医学院青年科技启动基金(Y2011Z003)

摘　　要：目的：建立大鼠成骨细胞的分离、培养方法,并进行改良.方法：分别应用酶消化法、传统的植块法和采用胶原酶消化结合骨片贴附法进行成骨细胞原代培养,通过细胞形态学、Von Kossa钙结节染色和免疫印迹对细胞纯度进行鉴定,通过生长曲线对3种方法取得的原代细胞生长情况进行比较.结果：3种培养方式均可获得高纯度成骨细胞,应用胶原酶消化结合骨片贴附法可以长期获得具有稳定生物学特征的成骨细胞.结论：胶原酶消化结合骨片贴附法可以较长期获得高纯度成骨细胞.Objective： To establish and improve the methods of rat osteoblasts separation and culture. Methods： Enzyme digestion method, the traditional bone planting method and the collagenase digestion combined with bone-chip attachment method were used to culture the primary osteoblasts. The cell purity was determined by cell morphology, Von Kossa calcium nodules staining and Western blotting. Cell growth was identified by the growth curves. Results： With all these three culture methods, osteoblast of high purity was obstained. The eollagenase digestion combined with bone-chip attachment method could obtain osteoblasts with stable biological characteristics in the long term. Conclusion： The collagenase digestion combined with bone-chip attachment method can obtain osteoblasts with high-purity for a relatively long time.

关 键 词：成骨细胞 原代培养 胶原酶消化法 骨片贴附法 

分 类 号：Q813[生物学—生物工程] R68[医药卫生—骨科学]