In vitro isolation and cultivation of human axillary dermal papilla cells

Objective To investigate an efficient rapid method for the isolation and cultivation of human axillary dermal papilla cells. Methods Skin specimens with hair follicles were obtained from the axillary area of patients who received bromhidrosis surgery in the Department of Dermatology of the First Affiliated Hospital to Army Medical University from October 2015 to May 2016. The axillary dermal papilla cells were isolated by two-step enzyme digestion method, one-step digestion method and micro-dissection method separately. Then, axillary dermal papilla cells were cultured and identified. Differences in the operative procedure, separation efficiency and adhesion efficiency of dermal papilla cells, cell emigration duration, total operation duration and actual operation duration were compared among the above 3 methods. Results Compared with the one-step digestion method and micro-dissection method, the two-step enzyme digestion method showed simpler operative procedure, more than 30% separation rate and 96% adhesion rate of dermal papilla cells after 1 week. Moreover, the cell emigration duration was shortened by 3 - 4 days by the two-step enzyme digestion method. The two-step enzyme digestion method also showed longer total operation duration, but shorter actual operation duration compared with the one-step digestion method and micro-dissection method, as well as lower contamination rate compared with the micro-dissection method. Cultured axillary dermal papilla cells grew in an aggregative pattern in the early stage, but grew in a non-aggregative pattern after 6 passages. Immunofluorescence assay showed positive staining for laminin and collagen Ⅳ in axillary dermal papilla cells. Conclusion The modified two-step enzyme digestion method is a kind of simple, efficient and rapid method for the isolation of human axillary dermal papilla cells, and axillary dermal papilla cells can be harvested through this method by using a few specimens. Key words: Hair follicle; Cell separation; Cell culture techniques; Axilla; Dermal papilla cells