摘要
目的建立人蛛网膜细胞的体外培养方法,观察其体外生长的生物学特性。方法应用组织培养的方法,对人蛛网膜细胞进行体外原代和传代培养。观察培养的蛛网膜细胞体外生长的特征。并对培养细胞进行形态学和超微结构的检查和鉴定。结果人蛛网膜细胞可以在体外成功培养传代。原代培养细胞大多48 h贴壁,8 d后旺盛生长,15 d形成单层。传代培养细胞次日贴壁,10 d形成单层。培养细胞呈扁平多角形,细胞表面具有丰富的微绒毛。蛛网膜细胞胞质染色质丰富,分布均匀,胞浆中线粒体、粗面内质网中等发达,可观察到丰富的高尔基体、核糖体和少量溶酶体,中间丝和微丝形成细胞骨架。cytokeratin 8及18、人类白细胞抗原DR位点(HLA-DR)和胶质原纤维酸性蛋白(GFAP)单克隆抗体免疫荧光染色阳性。结论应用组织培养的方法可以成功获得原代和传代培养的人蛛网膜细胞,培养的细胞具有上皮细胞及神经胶质细胞的生物学特性。
Objective To probe a method of culturing arachnoid cells of human in vitro and observe its growth characteristics. Methods Arachnoid cells of human were planted in 25 cm^2 Falcon flask and the characteristics of cultured cells were observed by inverted phase contract microscope. The cultured cells were identified by immunofluorescence staining and transmission electron microscope. Results Arachnoid cells of human could be successfully cultured for many passages. Most of the cells attached to the plastic surface of flask during the first 48-hours＇cuhure. The cells grew prosperously at the 8 th day and formed a monolayer in 15 days. Passaged cells attached to the plastic surface of flask on the second day and formed a monolayer in 10 days. The monolayer was made up of flat, roughly polygonal cells. Abundance caryotin was uniformly distributed in the cytoplasm of arachnoid cells. Moderate quantity of rough endoplasmic reticulum and chondriosome were seen in the endochylema. Affluent Golgi＇s body, ribosome and small quantity of eytolysosome were also observed. Cytoskeleton was made up of intermediate filament and microfilament. The cultured cells were positive for cytokeratin 8 and 18, human leukocyte antigen DR （HLA-DR） and glial fibrillary acidic protein （GFAP）. Conclusion The arachnoid cells of human can be successfully cultured in vitro and the cultured cells have some characteristics similar to the epithelium and glial cells.
出处
《中华神经外科疾病研究杂志》
CAS
2008年第3期245-248,共4页
Chinese Journal of Neurosurgical Disease Research
