人眼脉络膜黑色素细胞的体外培养及透射电镜观察  

In vitro culture and ultrastructure of human choroidal melanocytes

作　　者：王玲[1] 卢清君[2] 王宁利[2] 

机构地区：[1]青岛大学医学院附属医院,山东青岛266003 [2]首都医科大学附属北京同仁医院

出　　处：《山东医药》2007年第12期1-2,共2页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(30400229)

摘　　要：目的建立人眼脉络膜黑色素细胞体外培养及其超微结构研究方法。方法采用胰蛋白酶与胶原酶多次消化法对人眼脉络膜黑色素细胞进行取材和培养,利用光镜和透射电镜对培养细胞进行观察。结果初分离的脉络膜黑色素细胞呈棕色球状体,24 h内贴壁。贴壁细胞渐变为双极、三极或树枝状,胞质内含有棕黄色色素颗粒,传代后色素量保持稳定。电镜下可见细胞表面有少量微绒毛,胞质内黑素体沿胞膜分布或呈簇状分布。结论胰蛋白酶与胶原酶多次消化并在培养液中添加多种刺激因子,可建立人眼脉络膜黑色素细胞的有限细胞系。[Objective] To establish the culture of human choroidal melanocytes and observe their ultrastructure. [Methods] Cboroidal melanocytes were isolated from 4 eyeballs of 4 donors by trypsin-collagenase digestion. Isolated cells were cultured with F12 medium supplemented with fetal bovine serum,basic fibroblast growth factor,isobutyl methylxanthine and cholera toxin. Light and transmission electron microscopes were used to observe the characteristics of human choroidal melanocytes. [Results] Separated choroidal melanocytes showed brown spheropiasts and attached in 24. Attached cells displayed dipolar,tripolar or dentritic morphology. Yellowbrown pigment granules were seen in the cytoplasm, and melanin content remained stable after several passages. Ultrastructurally,sparing microvillus existed on the surface of cells,and melanosomes in the cytoplasm exhibited along membrane or in clump. [Conclusion] Trypsin-collagenase digestion and medium supplemented with special factors are helpful to establish a human choroidal melanocyte line.

关 键 词：细胞培养 脉络膜 黑色素细胞 透射电镜 

分 类 号：R773.4[医药卫生—眼科]