人胎儿雪旺氏细胞的体外培养及其纯化研究  

CHARACTERIZATION AND PURIFICATION OF HUMAN FETAL SCHWANN CELLS IN VITRO

作　　者：严恒林[1] 沈馨亚[1] 刘才栋[1] 陈丽琏[1] 汪洋[1] 王自美[1] 

机构地区：[1]上海医科大学解剖学教研室,上海脑研究所

出　　处：《神经解剖学杂志》1994年第4期289-293,共5页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金

摘　　要：本实验用酶消化法从孕１４～１９周人胎儿周围神经中分离培养雪旺氏细胞，结合差速贴壁和阿糖胞苷处理进行纯化，根据Ｓ－１００蛋白免疫细胞化学染色和形态学特征，分析不同时期雪旺氏细胞的纯化程度。实验结果显示：培养４ｄ，倒置相差显微镜下观察，雪旺氏细胞呈典型的双极或三极形，端对端或旋涡状排列，Ｓ－１００蛋白免疫细胞化学染色呈阳性反应，其比例约占９８％；在２～３周内，雪旺氏细胞的纯度维持在８５％～９０％；培养３５ｄ，雪旺氏细胞约占８０％；单纯采用阿糖胞苷或差速贴壁处理，培养３５ｄ，雪旺氏细胞约占７０％；原代培养９２ｄ，传代１１代细胞生长良好。实验结果表明，多种方法联合使用较单一方法所得的纯度高。本方法快速、简便，能为雪旺氏细胞作为移植材料提供足够的来源。Highly purified population of Schwann cells (SC)from human fetal nerves has been well prepared and desczibed. Cultures were prepared by chemical and mechanical dissociation and purification were developed in combining differential adhesion and two pulsed cytosine arabinoside (Ara-C) treatment.A time course analysis of Schwann cell marks at different time in vitro was performed employing immunoperoxidase to determine the percentage of SC in culture and evaluate the maintenance of specific SC.Approximately 97 %～98% of cells were typical bipolar and S-100 protein positive in 4 days in vitro (DIV), about 85%～90% in 14～18 DIV according to cytological and immunocytochemical criteria. In 35 DIV, 80% were SC while less than 70% of SC in control groups by either differential adhesion or Ara-C treatment only. The primary and its up to 11 passages of SC grew very well even in 92 DIV. Further observation of the SC obtained by this method were also taken under both SEM and TEM in displying its typical morphological charaererizations.

关 键 词：雪旺氏细胞 细胞培养 人胎儿 提纯 

分 类 号：R329－33[医药卫生—人体解剖和组织胚胎学]