组织工程化许旺细胞培养及纯化的实验研究  

A Study of Different Methods to Cultivate and Purify SCs in Vitro

作　　者：田宏斌[1] 周梁[1] 莫晓芬[1] 田洁[1] 

机构地区：[1]复旦大学附属眼耳鼻喉科医院,上海200031

出　　处：《中国眼耳鼻喉科杂志》2005年第4期220-221,i003,共3页Chinese Journal of Ophthalmology and Otorhinolaryngology

基　　金：上海市科委基金资助(020J14026)

摘　　要：目的获得组织工程化的乳鼠许旺细胞.方法采用出生1 d乳鼠的脊髓后根冲经节,以复合酶消化法进行乳鼠许旺细胞培养.通过Sigma-cote处理滴管和加入神经生长因子减少细胞损失、加速细胞分裂、增加细胞总量;通过低浓度胰酶快速消化、差速贴壁法的有机结合对许旺细胞进行纯化;继用抗S-100蛋白单抗通过(即用型)ABC免疫组化检测试剂盒,SABC-CY3试剂盒进行细胞鉴定.结果细胞纯度＞90%,细胞量＞2×10 7/ml.结论本实验方法具有高度重复性,可获得高纯度,大量的许旺细胞.Purpose To develop a method to culture Swchwann cells for tissue engineering.Methods One-day-old rats were sacrificed by decapitation and their Dorsal Root Ganglions were removed for primary culture.By adding nerve growth factor in dish and Sigma-cote painting burettes,quality of Schwann cells can be improved.By rapid low concentration trypsinization and differential adhesion,purity of ceils can also be improved.The cells were identified with S-100 protein antibody by ABC methods and SABC-CY3 methods.Results The purity of ceils was more than 90% and the quality was more than 2×10~7/ml.Conclusion These methods to cultivate Swchwann cells for tissue engineering were repeatable,high purity and quality cells can be harvestd.

关 键 词：组织工程化 细胞培养 实验研究 纯化 ABC免疫组化 S-100蛋白 许旺细胞 神经生长因子 后根神经节 Sigma 检测试剂盒 酶消化法 细胞分裂 有机结合 细胞鉴定 实验方法 乳鼠 细胞损 低浓度 贴壁法 即用型 细胞量 重复性 高纯度 

分 类 号：R318.0[医药卫生—生物医学工程]