小鼠颅底软骨联合细胞的体外原代培养  

The Primary Culture of Chondrocytes of Mouse Cranial Base Synchondrosis

作　　者：高国杰[1] 沈绍莹[1] 胡江天[1] GAO Guo-jie;SHEN Shao-ying;HU Jiang-tian(Dept. of Orthodontics,School of Stomatology,Kunming Medical University,Kunming Yunnan 650031,China)

机构地区：[1]昆明医科大学口腔医学院正畸科,云南昆明650031

出　　处：《昆明医科大学学报》2017年第7期37-40,共4页Journal of Kunming Medical University

基　　金：云南省教育厅科学研究基金资助项目(2017ZZX203);昆明医科大学科技创新团队建设项目(CXTD201607)

摘　　要：目的探讨小鼠颅底软骨联合细胞体外原代培养的方法并了解其基本生物学特征.方法显微手术采集小鼠颅底蝶枕软骨联合组织;用胰蛋白酶和胶原酶联合消化软骨基质,获取软骨细胞;用含血清的DMEM培养基进行原代和传代细胞培养;通过显微光学成像、生长曲线描记及免疫组织化学染色观测细胞形态、活力及II型胶原合成能力.结果培养细胞在一定传代次数内稳定保持软骨细胞的典型形态和功能特征.结论手术分离配合酶联合消化是小鼠颅底软骨联合细胞体外原代培养实验的一条可靠途径.Objectives To explore the experimental approach for in vitro primary culture of chondrocytes of mouse cranial base synchondrosis and to investigate its basic biological features.Methods The spheno-occipital synchondrosis of new natal mouse was harvested through surgical dissection,from which chondrocytes were further isolated through enzymatic digestion with trypsin and type II collagenase.Chondrocytes of the primary and successive generations were cultured with serum containing Dulbeccomodified eagle's medium(DMEM).Optical microscopic imaging,growth curve tracing and immunohistochemistry were conducted to observe the morphological features,growth behavior and type II collagen expression.Results Within five generations,the cultured cells consistently presented typical morphological and functional features of chondrocyte.Conclusion The surgical tissue dissection combined with double enzymatic digestion is a reliable experimental approach for the primary culture of chondrocytes of mouse cranial base snychondrosis.

关 键 词：小鼠 颅底 软骨联合 软骨细胞 细胞培养 

分 类 号：R813.11[医药卫生—放射医学]