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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 investigate the feasibility of culture yellow choroidal and iris melanocytes in vitro and to observe whether melanocytes can be synthesized in vitro. Methods The choroid and iris stromal tissue were removed by trypsin and collagenase. The cells were separated and cultured with F12 medium supplemented with fetal calf serum, basic fibroblast growth factor, isobutylmethylxanthine and cholera toxin to cultivate. Immunohistochemical staining with keratin antibody and S 10 0 antibody. The intracellular melanin content was measured with a spectrophotometer and its production was calculated. Results The uveal melanocytes were successfully cultured in vitro. The average pigment content per melanocyte in the growing period was (79 83 ± 36 2 0) pg, and melanin (1144 ± 5 77) pg per day was synthesized in vitro. Of uveal melanocytes were positive for S 10 0 but negative for keratin. Conclusion The yellow human uveal melanocytes cultured in vitro can synthesize pigment in vitro. The pigment content of yellow uveal uveal melanocytes is between that of white race and black race.