毛囊无黑素性黑素细胞的分离和培养

目的：分离和培养毛囊无黑素性黑素细胞，探讨其生物学特性。方法：采用胶原酶V游离毛囊，0．5％分离酶消化婴幼儿包皮。0．05％胰酶和0．53mmol／L乙二胺四乙酸(EDTA)消化游离的毛囊和来自包皮的表皮，分别制备单细胞悬液进行培养。传至第3代，待细胞爬上盖玻片后，分别以NKI／heteb和HMB-45单抗染色。结果：毛囊的培养细胞，绝大多数为双极细胞，增殖较快，被NKI／beteb单抗识别，而HMB-45染色阴性，包皮的黑素细胞染色结果则与之相反．结论：毛囊的外根鞘中存在功能和形态上不成熟的无黑素性黑素细胞(AMMC)、AMMC长期培养的成功，有利于研究AMMC在毛囊生物学中所起的作用。

Isolation and culture of human hair-follicle amelanotic melanocytes

Objective:To isolate and culture melanocytes derived from human hair follicles, and investigate their characteristics. Methods: Hair follicles of normal human scalp were isolated by collagenase V treatment. The epidermis taken from children's foreskin was treated with 0.5% dispase. The cell suspensions from the hair-follicle and the epidermis were prepared by trypsin/ethylenediamine tetraacetic acid treatment and cultured respectively in a mixture of Eagle's minimum essential medium (supplemented with 12-O-tetradecanoyl-phorbol-13-acetate and cholera toxin) and keratinocyte serum-free medium. In the third passage ,the cells were confirmed by staining with NKI/beteb and HMB-45 monoclonal antibodies respectively. Results : After the contaminating keratinocytes were removed, the cultured cells from hair-follicle with two distinct morphologies remained, the most of cells were small bipolar, which proliferated very rapidly and could be recognized by NKI/beteb monoclonal antibody, not recognized by HMB-45 monolonal antibody. Conclusions: Amelanotic melanocytes located in the outer root sheath (ORS) are immature morphologically and functionally. The availability of cultured hair-follicle melanocytes will facilitate investigations of the role of these cells in normal and abnormal hair biology.