[蒙古人葡萄膜黑色素细胞体外黑色素生成的培养与研究]

[Culture and study on melanogenesis of mongolian uveal melanocytes in vitro].

作者信息

Zhou Xiang-tian, Qu Jia, Fu Xiao-ying, Lu Fan, Hu Dan-ning

机构信息

School of Optometry and Ophthalmology, Wenzhou Medical College, Wenzhou 325000, China.

出版信息

Zhonghua Yan Ke Za Zhi. 2004 Feb;40(2):104-7.

PMID:15059563

Abstract

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.

摘要

目的

从成年供体眼中建立蒙古人葡萄膜黑色素细胞系，并研究这些细胞在体外的黑色素生成。

方法

去除色素上皮后，通过胰蛋白酶-胶原酶消化分离葡萄膜黑色素细胞。将分离的细胞接种于添加胎牛血清、碱性成纤维细胞生长因子、异丁基甲基黄嘌呤和霍乱毒素的F12培养基中培养。用分光光度计测量黑色素含量。通过公式计算黑色素生成量。免疫细胞化学研究中使用的抗体为抗细胞角蛋白抗体和抗S-100抗体。

结果

通过该方法获得了纯黑色素细胞培养物。培养物中的所有细胞对S-100抗体呈阳性染色，但对细胞角蛋白抗体不染色，表明它们是葡萄膜黑色素细胞的纯培养物。在生长的葡萄膜黑色素细胞中，黑色素含量为（79.83±36.20）pg/细胞（平均值±标准差）。黑色素生成量为（11.44±5.77）pg/细胞/24小时。