人眼小梁细胞体外培养及生物学特性研究

The Study of the Biological Character on the Culture of Human Trabecular Meshwork Cells in Vitro

目的:建立人眼小梁细胞培养方法并研究其生物学特性。方法:以体外组织块培养的方法获得培养的人小梁细胞,应用光镜、电镜观察细胞的形态学特征,并观察其免疫组化特性和细胞的生长曲线。结果:光镜下小梁细胞为扁平多角形、单层生长;电镜下细胞连接为点粘连和缝隙连接、细胞表面可见微绒毛、胞浆细胞器丰富;免疫组化染色对抗纤维连接蛋白(Anti—FN)、抗层粘连蛋白(Anti—LN)、抗神经元特异性烯醇化酶(Anti—NSE)单抗呈阳性,对抗第Ⅷ因子(Anti-ⅧFactor)单抗呈阴性;传代小梁细胞繁殖时间较长,10天后为平台期。结论:根据体外培养的小梁细胞的形态特点、生长特征、免疫组化特性可对其进行鉴定。人小梁细胞体外培养的成功,为在细胞和分子水平研究青光眼的发病机制提供了有利的条件。眼科学报 1996;12:64—69。

Purpose: To establish the culture of human trabecular cells in vitro and to study their biological character. Method: The trabecular specimens from human eyes were cultured. The morphologic features of cultured cells were examined by light and electron microscopy, the immunohistochemical characteristics and the proliferative curve of the cultured cells were also investigated. Results: The primary cells in culture are multiangular or irregular, and the confluent trabecular cells grow as a single cell layer. The junctions between trabecular cells observed most frequently are puncta adherens and gap junction. Trabecular cells showed apical villons projections and had a high density of various organelles. The trabecular cells were stained intensely with monocolone antibodies to fibronection and laminin and NSE. And it had negative reaction to monocolone antibodies to VI factor. The cultured cells showed a longer doubling time. Conclusion: The culture of human trabecular cells in vitro was established. This was a key step to dissect the trabecular specimen accurately and carefully. According to the different celluar growth patterns and speed, morphologic and immunohistochemical characteristics from that of the near cells, the cultured human trabecular cells can been identified. Eye Science 1996; 12: 64- 69.