P43 SERIAL CULTIVATION OF HUMAN DERMAL PAPILLA CELLS RETAINING THE HAIR-INDUCING ABILITY

¹Koh-éi Toyoshima*, ¹Yoshie Sasaki, ²Akio Satoh, ³Mutsumi Inamatsu, ³Souichi Oomizu, ^3,4Katsutoshi Yoshizato; ¹Bioart Laboratory, ²Tokyo Memorial Clinic, ³Hiroshima Tissue Regeneration Project Joint-Research Project for Regional Intensive, JST, ⁴Dept. of Biological Science, Graduate School of Science, Hiroshima University

Human dermal papilla cells show a lower proliferative capacity than dermal fibroblasts in culture, and lose their inherent functions to induce hair follicles in the epidermis when cultured at more than 4-5 passage. Inamatsu et al. established serial cultivation of rat papilla cells by maintaining them with the conditioned medium of keratinocytes and FGF. These cells sustained their hair-inductive ability during the culture for more than 70 passages. We applied this culture method to the human papilla cells. Dermal papillae were isolated from human scalp skin and these cells were cultured in 3 conditions: (1) Dulbecco  Modified Eagle Medium with 10% fetal bovine serum (DMEM10), (2) DMEM10 containing 5 ng / ml FGF2, and (3) DMEM10 containing 50% conditioned medium of keratinocytes derived from rat sole skin (CM). We found that CM-containing medium was the most effective in inducing cell proliferation and increasing serial passage. During subcultivation of human dermal papilla cells, the growth-stimulatory effects of CM was decreased. However, we could culture them until at passage number of 10-16. The cultured cells were found to retain the innate hair follicle inductive ability at until at least 11 passages. The culture method developed in this study should facilitate the research on the characterization of human dermal cells.