体外培养的牛眼小梁细胞及微管微丝的形态学观察

Cultivation of bovine trabecular meshwork cells in vitro and morphological observation of their microtubules and microfilaments

目的 :初步观察体外培养的牛眼小梁细胞 (tra becularmeshworkcells,TMcell)及微管微丝的形态 ,为进一步探讨原发性开角型青光眼 (primaryopen angleglaucoma,POAG)的发病机制提供理论依据。方法 :①TM的体外培养 ,收稿日期 :2 0 0 1-11-19;修回日期 :2 0 0 2 -0 3 -2 0作者简介 :沈凤梅 (1964 -) ,女 ,山东人 ,医学硕士。通信作者 :沈凤梅 (E -mail:wanglgu @pub .xaonline .com)。应用免疫组化方法 (神经原特异性烯醇化酶 ,Ⅷ因子相关抗原染色 )进行细胞鉴定。用光学及透射电子显微镜对细胞进行形态学及生长特性的观察。②免疫组化法肌动蛋白(actin)、微管蛋白 α(tubulin α)染色。结果 :①TM细胞培养成功 ,以上皮型为主。②小梁细胞内富含微丝 ,与细胞长轴平行 ,并密集于细胞的周边部 ;微管以核周为主。结论 :牛眼小梁细胞体外培养技术是研究小梁细胞特性的重要实验技术。小梁细胞富含微管微丝 。

Objective:The purpose of this investigation was to cultivate bovine trabecular meshwork cells in vitro and observe their microtublin and microfilament forms,in an attempt to learn about the pathogenesis of primary open angle glaucoma(POAG).Methods:①Bovine trabecular meshwork cells were cultured in vitro:Fresh bovine eyeballs were collected. The tissues were cultured primarily and the cells subcultured. The morphological features and ultrastructure of the bovine trabecular meshwork cells were studied under light microscopy and transmission electron microscopy.②Histological identification:The bovine trabecular meshwork cells were identified by the immunohistochemical strept avidin biotin complex method with neuronal specific enolase(NSE) and factor Ⅷ:Ag related antigen(Ⅷ:Ag).③Actin and tublin α were stained by immunohistochemistry methods.Results:①Morphological features:Primary bovine trabecular meshwork cells appeared in round,fusiform and unregulated shape with several cell processes. The confluent cells showed as an epithelial monolayer. Cultured bovine trabecular meshwork cells have cytoplasm,organelles and granum in abundance. They also have a double nucleus membrane and obvious nucleoles.②The immunohistochemical stains showed that the bovine trabecular meshwork cells presented positive reactions to NSE and negative reaction to Ⅷ:Ag,which proved that bovine trabecular meshwork cells are derived from embryonic neural crest.③Dense bundles of microfilament were located parallel to the long axis of the cell,primarily beneath the surface of the cells. Numerous microtubules gathered around the nucleus.Conclusion:①Bovine trabecular tissue is abundant and its anatomic boundary line is distinct. Establishing a method for culturing bovine trabecular meshwork cells is important for investigating the characteristics of these cells.②Trabecular meshwork cells have abundant microfilaments and microtubules and two major cytoskeletal elements,which play an important role in structural integrity and motile function in the trabecular meshwork.