人眼筛板细胞体外培养的研究

Cell culture of human lamina cribrosa in vitro

目的研究筛板细胞合成细胞外基质的生理与病理，建立筛板细胞培养模型。方法采用酶消化法对１０只人胚眼筛板组织进行体外培养，并利用电镜和免疫组化技术鉴定。结果所培养的细胞呈多角扁平状，核周胞浆内富含颗粒，核相对透明，核仁清楚；细胞表面有微绒毛，核膜周围有异染色质，胞膜附有基底膜及细胞外基质；免疫荧光染色对Ⅰ、Ⅲ、Ⅳ型胶原及纤维连接蛋白，层粘连蛋白呈阳性；对神经胶质纤维酸性蛋白，Ⅷ因子相关抗原呈阴性。以上均符合筛板细胞的特点，此细胞株现已传代至第４代。结论成功地建立筛板细胞体外培养模型，可用于青光眼发病机制的研究。

Objective To investigate the physiology and pathology of the biosynthesis of extracellular matrix (ECM) by lamina cribrosa cells (LCC) and to establish a model of cultured LCC. Methods The LCC of 10 human dead fetal eyes were cultured with enzyme digestion procedures in vitro. The cultured cells were identified by immunohistochemistry assays and electron microscopy. Results The cultured cells were flat, polygonal in shape, and their cytoplasm contained abundant perinuclear granules. The nuclei were relatively transparent and nucleoli were clear, there were basement membrane like material and ECM surrounding the cell membrane. The immunofluorescent staining was positive for collagen type Ⅰ,Ⅲ,Ⅳ, fibronectin and laminin, negative for glial fibrillar acid protein and factor Ⅷ. The characteristics mentioned above were coincident with the morphology of LCC. This cell line has been cultured to its fourth generation. Conclusion We have successfully cultured a cell line of LCC, and it may be utilized in the study of pathogenesis of glaucoma.