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目的观察不同培养条件及时相对体外培养的正常人黑素细胞生物学特性的影响。方法分离正常成人包皮黑素细胞,分别在添加有碱性成纤维细胞生长因子(bFGF)和霍乱毒素(CT)的RPMI1640培养基及角质形成细胞条件培养基中培养。培养的细胞用L-Dopa染色及S-100蛋白染色鉴定。观察其形态、增殖特性。结果培养细胞经生物学鉴定为黑素细胞。原代培养及条件培养基传代培养中黑素细胞有多个突起,而bFGF/CT培养基中传代培养时黑素细胞多呈梭形。两种培养条件下黑素细胞增殖特性无明显区别。结论两种方法均可稳定获得黑素细胞的体外选择性纯培养,角质形成细胞可通过细胞间接触及分泌细胞外因子影响黑素细胞的树突形成。
Objective To observe the effects of different culture conditions on the biological characteristics of normal human melanocytes cultured in vitro. Methods Normal human preputial melanocytes were isolated and cultured in RPMI 1640 medium supplemented with basic fibroblast growth factor (bFGF) and cholera toxin (CT) and keratinocyte conditioned medium. The cultured cells were identified by L-Dopa staining and S-100 protein staining. Observation of its morphology, proliferation characteristics. Results Cultured cells were biologically identified as melanocytes. In primary culture and conditioned medium, there are many melanocytes in the subculture, while in the bFGF / CT medium, the melanocytes are mostly fusiform. There was no significant difference in melatonin proliferation between the two culture conditions. Conclusion Both methods can obtain melanocytes in vitro selectively and selectively. Keratinocytes can affect the dendrites formation of melanocytes by intercellular contact and secretion of extracellular factors.