人表皮黑素细胞、毛囊无色素黑素细胞和鼠黑素瘤细胞的原子力显微镜观察

Morphologic comparison of cultured human epidermis melanocytes,amelanotic melanocytes from hair follicles and mouse melanoma cells observed by atomic force microscope

目的:观察培养的人表皮黑素细胞、毛囊无色素黑素细胞和S91鼠黑素瘤细胞的形态结构。方法:培养并纯化来自正常人包皮的黑素细胞以及来自毛囊的无色素黑素细胞,同时复苏S91细胞株,传代后接种到内置云母片的培养板中,细胞贴附到云母片上后固定,用原子力显微镜扫描观察。结果:正常人表皮黑素细胞有3级分支,在主干及分支的顶端和侧缘可见膨出的球形结构。鼠黑素瘤细胞仅有很短的2级分支,在2级树突近端可见黑素小体。毛囊无色素黑素细胞只有1级树突,并只在树突近端有少数黑素小体。结论:表皮黑素细胞在形态上比黑素瘤细胞、毛囊无色素黑素细胞更成熟,有更多的黑素颗粒从树突的顶端和侧缘以胞吐的形式被输出。

Objective:To image the surface structure of cul tured human epidermis melanocytes,hair follicle amelanotic melanocytes and mouse melanoma cells by atomic force mi cro scope.Methods:Human hair follicles were isolated by colla-genase V treatment.The epidermis obtained from children's fore skin was treated with0.5%dispase.The cell suspensions from hair-follicle and epidermis were prepared respectively and seeded in six-well plates,in which sheets of mica were placed.The mouse melanoma cells were passaged from S91cell line.Sample preparation for AFM was fixed on mica and AFM im ages of scanning observation were cap tured by contacting and tapping modes and operated under normal atmo spheric pressure and temperature.Results:Human epidermal melanocytes had three grades of branches.Globular structures protrud-ed on the tops and lateral sides of main branches and subordinate branches.There were many250～620nm diame ter gran-ules on the dendrite surface of epidermal melanocytes.There were two grades of branches and a few granules in S91mouse melanoma cells.Most cultured amelanotic melanocytes from human hair follicles were bipolar in shape.Conclutions :The second--third grade main branches and subordinate branches might take part in transferring melanosomes into the ker-atinocyte s in addition to transferring melanosome through the tops of dendrites.The melanin gran ules can be ex pelled by ex-ocytosis.