成纤维细胞生长因子对黑素细胞生长的影响

Effects of basic fibroblast growth factor （bFGF） on the growth of melanocytes

目的 探讨成纤维细胞生长因子（bFGF）对体外培养正常人黑素细胞增殖的影响,寻找快速体外培养正常人黑素细胞的方法.方法 分别用普通的合成培养基与添加一定浓度bFGF的合成培养基原代培养正常人黑素细胞,观察两种条件下细胞生长情况,并对培养的黑素细胞进行鉴定,对不同生长期黑素细胞的形态进行观察,测定不同浓度bFGF（0.3、0.6、0.9、1.2、1.5、1.8、2.1 μg/L）对黑素细胞增殖、酪氨酸酶活性的影响.结果 采用培养基中加入一定浓度bFGF的方法成功培养出免疫组化染色阳性的纯化黑素细胞,该细胞不同生长期形态有一定差异.bFGF处理组黑素细胞生长较快,培养基中加入0.3、0.6μg/L bFGF时黑素细胞的增殖率显著高于对照组（P＜0.05或P＜0.01）,加入1.5、1.8μg/L bFGF时酪氨酸酶活性显著高于对照组（P＜0.05或P＜0.01）.结论 在合成培养基中加入一定浓度的bFGF能缩短原代培养时间,短期内可获得大量生长活力好的纯化黑素细胞.一定浓度的bFGF能促进黑素细胞增殖和酪氨酸酶活性.

Objective To study the effects of bFGF on the proliferation of cultured human melanocytes, and to seek a quick method for in vitro culture of human melanocytes. Methods Melanocytes were isolated from human foreskin, and divided into two parts to be cultured with or without the presence of bFGF （0.3 μg/L）. Second-passage melanocytes were identified with immunochemical stain. The growth of melanocytes was observed every 3 days for 12 days. Third-passage melanocytes were treated with various concentrations （0.3 - 2.1 μg/L） of bFGF for 72 hours followed by the detection of proliferation of and trosinase activity in melanocytes. Results Human melanocytes were obtained from primary culture in medium containing certain concentrations of bFGF, which were identified with immunohistochemical stain. The morphology of cultured melanocytes varied with growth stage of cells. The bFGF-treated melanocytes appeared to grow more rapidly than untreated melanocytes. Further more, a significant increase was observed in the proliferation rate of melanocytes treated with bFGF of 0.3 and 0.6 μg/L （P〈0.05 or 0.01 ） and tyrosinase activity in melanocytes treated with bFGF of 1.5 and 1.8 μg/L （P 〈 0.05 or 0.01 ） in comparison with the untreated melanocytes.Conclusions The addition of certain concentrations of bFGF to defined medium can benefit the primary culture of melanocytes and make it possible to get large quantities of purified melanocytes with high viability in short periods. Certain concentrations of bFGF can up-regulate the proliferation of and tyrosinase activity in melanocytes.