【摘要】 旨在探讨羊驼皮肤黑素细胞体外培养和鉴定的方法,为建立羊驼皮肤黑素细胞系奠定基础,也为毛色基因功能和体细胞克隆等研究提供理想的生物材料。采用DispaseⅡ酶和胰酶/EDTA两步消化法获取羊驼皮肤黑素细胞,并进行体外原代和传代培养。观察细胞各个阶段的形态,通过生长曲线等手段研究羊驼皮肤黑素细胞的增殖能力,利用多巴(Dopa)染色、免疫细胞化学染色、电镜、及RT-PCR等方法对所获得的细胞进行鉴定。在生长培养基作用下,细胞增殖旺盛;多巴染色、电镜、RT-PCR及免疫组化染色结果均表明培养黑素细胞的生物学功能正常,保持了正常的生物学特性。本研究体外成功建立了羊驼皮肤黑素细胞培养体系。更多还原

【Abstract】 The aim of this study was to explore the method for culture and identification in vitro of the alpaca skin melanocytes,to lay a foundation for the establishment of the alpaca skin melanocytes lines,and to provide an ideal biological material for coat color genes function and somatic clone studies.Alpaca skin melanocytes were obtained by the two step digestion method of DispaseⅡ and trypsin/EDTA,and the primary and secondary culture in vitro were carried out.Morphological observation and growth curve were used to evaluate the proliferation and differentiation ability of alpaca skin melanocytes.The cells were identified with Dopa-staining,immunochemical staining,electron microscope and RT-PCR.The cells showed strong proliferation ability in the proliferative media.Dopa-staining showed that the cultured alpaca skin melanocytes had melanin synthesis,immunochemical staining,RT-PCR and electron-microscopy technology assay demonstrated that the cell differentiation was normal.The results showed that the culture system was established successfully in vitro.更多还原