摘要
目的 探讨体外培养黄色人种脉络膜、虹膜黑色素细胞的可行性 ,并观察黑色素细胞能否在体外培养时合成色素。方法 将除去色素上皮细胞的脉络膜、虹膜基质组织用胰酶与胶原酶消化 ,分离得到的细胞用添加胎牛血清、碱性成纤维细胞生长因子、异丁基甲基黄嘌呤、霍乱毒素的F12培养液培养。用角蛋白抗体、S 10 0抗体进行细胞免疫组化染色。用分光光度计测定细胞内黑色素含量并计算其生成量。结果 成功体外培养了葡萄膜黑色素细胞 ;生长期每个黑色素细胞内的平均色素含量为 (79 83± 36 2 0 ) pg ,每个细胞每天合成黑色素 (11 4 4± 5 77) pg ,体外培养的葡萄膜黑色素细胞S 10 0表达阳性 ,但角蛋白表达阴性。结论 体外培养的黄色人种葡萄膜黑色素细胞能在体外合成色素 ,黄色人种葡萄膜黑色素细胞内的色素含量与制造量介于白色人种和黑色人种之间。
Objective To establish cell lines of Mongolian uveal melanocytes from adult donor eyes, and to study melanogenesis of these cells in vitro. Methods After removal of pigment epithelium, UM were isolated by trypsin-collagenase digestion. Isolated cells were cultured with F12 medium supplemented with fetal bovine serum, basic fibroblast growth factor, isobutylmethylxanthine and cholera toxin. Melanin content was measured by spectrophotometer. Melanin production was calculated by a formula. The antibodies used in immunocytochemical studies were anti-cytokeratin and anti-S-100 antibodies. Result Pure melanocytes cultures were obtained by this method. All of the cells in the cultures stained positively with antibodies to S-100, but not cytokeratin, indicating that they were pure culture of UM. In the growing UM, melanin content was (79.83±36.20) pg/cell (mean±SD). Melanin production was (11.44±5.77) pg per cell/24 h. Conclusion Uveal melanocytes from Mongolian eyes are successfully cultured. Cultured uveal melanocytes can synthesize melanin in vitro. Melanin content and melanin production of Mongolian UM lie between those from black and Caucasian eyes.
出处
《中华眼科杂志》
CAS
CSCD
北大核心
2004年第2期104-107,共4页
Chinese Journal of Ophthalmology
基金
国家自然科学基金资助项目(30371507)
浙江省自然科学基金青年人才基金资助项目(RC9606)
浙江省自然科学基金资助项目(302095)
浙江省自然科学基金资助项目(ZB0202)
浙江省卫生厅重大课题基金资助项目(2002ZD009)
浙江省科技厅科技攻关计划-重点科研项目-社
