人体皮肤Merkel细胞原代培养与鉴定

Primary culture and identification of human skin Merkel cells

目的探索人体皮肤Merkel细胞体外培养方法,为进一步研究银屑病发病中Merkel细胞的神经内分泌调节作用积累基础资料。方法获取正常人皮肤组织,采用酶消化法分离细胞。设计专门的细胞体外培养系统,观察Merkel细胞在3种不同培养液中贴壁和增殖情况。采用Merkel细胞特异性标记抗体CK20标记Merkel细胞,采用免疫组化和流式细胞仪进行鉴定。结果人体皮肤Merkel细胞在3种培养液中均能顺利贴壁和增殖,并具有内分泌功能,但1号培养液[Ham's-F12+胎牛血清(10/100 ml)+b FGF(20 ng/ml)]更有利于Merkel细胞生长。经流式细胞仪鉴定,Merkel细胞纯度可达90.11%。结论在本培养系统中,人体皮肤Merkel细胞能成功进行体外培养,并可获得较高纯度的Merkel细胞。

Objective To explore the culturing method for human skin Merkel cells in vitro and study the adjustment effects of Merkel cells on neuroendocrine function in the pathogenesis of psoriasis. Methods Normal human skin tissues were obtained,and the cells were isolated by enzymatic digestion method. The adhesion and proliferation of Merkel cells in three different culture media were observed by means of a special designed in-vitro cell culturing system. Specific Merkel cell marker antibody CK20 was used to label Merkel cells,and immunohistochemistrical method and flow cytometry were used for cell identification. Results The human skin Merkel cells could smoothly complete adhesion and proliferation in three culture media,and all had an endocrine function,but No. 1 culture medium（Ham ˊs-F12＋ fetal bovine serum（10/100 ml）＋bFGF（20 ng/ml）was more conducive to the growth of Merkel cells. According to the identification by the flow cytometer,the purity of Merkel cells could be up to 90. 11%. Conclusion In this culture＆amp;nbsp;system,human skin Merkel cells can be successfully cultured in vitro,and high-purity Merkel cells can be obtained.