大鼠少突胶质前体细胞在化学条件培养基中定向分化的形态学观察  

Morphological observation of directional differentiation from oligodendrocyte precursors to mature oligodendrocyte by chemical conditioned medium in vitro

作　　者：付元山[1] 王滨[1] 汪洋[2] 刘才栋[2] 沈馨亚[2] 

机构地区：[1]大连医科大学解剖学教研室,辽宁省大连市116027 [2]复旦大学上海医学院解剖学教研室,上海市200032

出　　处：《中国临床康复》2006年第1期15-17,共3页Chinese Journal of Clinical Rehabilitation

摘　　要：目的:观察体外无血清化学培养条件下少突胶质前体细胞向成熟少突胶质细胞的定向分化。方法:实验于2000-04/07在复旦大学上海医学院解剖学教研室完成。①取新生SD大鼠获取少突胶质前体细胞;前O2A祖细胞原代混合培养7d后,以B104神经胶质瘤细胞株条件培养液犤100mL含:牛血清白蛋白每20mg,胰岛素2.5mg,转铁蛋白20mg,腐胺1.6mg,亚硒酸钠0.5μg,黄体酮0.6μg,生物素0.24μg,T33μg,T40.04mg,溶于DMEM/F12对半配置的培养液中犦增殖并纯化的少突胶质前体细胞,然后将培养基更换为无血清的化学条件培养基。②倒置相差显微镜观察记录24,48和72h细胞的形态变化,分化成熟的少突胶质细胞作扫描电镜观察。结果:少突胶质前体细胞无血清化学条件培养基培养形态学演变:培养24h,少突胶质前体细胞突起增加为三极或四极,胞体增大稍迟;48h后突起明显增多,相互之间连接成网;72h,胞体大且边缘不规则,突起分支更加清晰,分支彼此呈三维立体交错,部分突起之间可见指环状或膜片状结构。结论:化学条件培养基可使在体外培养条件下的少突胶质前体细胞定向分化为成熟的少突胶质细胞,细胞形态呈现渐变的过程。AIM： To observe the differentiation to mature oligodcndrocyte from oligodendrocyte precursors in vitro by serum-free chemical conditioned medium. METHODS： The experiment was carried out in the Department of Anatomy, Shanghai Medical College of Fudan University between April and July 2000. ①The oligodendrocyte precursors were obtained from newborn SD rats, the oligodendrocyte precursors were proliferated and purified with the conditioned medium of B104 neuroglioma cell line （every 100 mL contained 20 mg bovine serum albumin, 2.5 mg insulin, 20 mg transferring, 1,6 mg putrescine, 0.5μg sodium selenite, 0.6 μg progesterune, 0.24 μg biotin, 3 μg T3, 0.04 mg T4, which were dissolved in the culture media of half-to-half DMEM/F12） after the oligodendrocytetype-2 astrocyte progenitors were primarily treated with mixed cultivation for 7 days. ②The morphological changes of the cells at 24, 48 and 72 hours were observed under inverted phase contrast microscope, and the differentiated mature oligodendrocytes were observed under scanning electron microscope, RESULTS： The morphological development of oligodendrocyte precursors cultured in serum-free chemical conditioned medium： The increase of apophysis of oligodendrocyte precursors at 24 hours was degree 3 or 4, and the cell body enlarged a little late; The apophysis obviously increased after 48 hours, and connected to net; The cell body was big with irregular margine at 72 hours, the branch of apophysis was much clearer, the branches were three-dimensionally crossed, and there were ring-like or membrane-like structures among part of the apophysis. CONCLUSION： The oligedendrocyte precursors could differentiate to mature oligodendrocyte by chemical conditioned medium in vitro and the cellular forms changed gradually.

关 键 词：细胞培养 少突神经胶质 细胞分化 大鼠 培养基 定向分化 形态学 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]