Establishment of rat dermal papilla cell lines that sustain the potency to induce hair follicles from afollicular skin

Dermal papilla cells in culture show a lower proliferative capacity compared with dermal fibroblasts, and lose their in situ potency to induce hair follicles in the epidermis at more than 10 passage numbers. This study overcomes these limitations of cultured papilla cells and for the first time demonstrates that papilla cells can be serially cultured for a long period without losing their hair-inductive potency. Outgrowth and the ensuing proliferation of papilla cells were markedly stimulated when explants of rat vibrissa papillae were cultured with rat sole-derived keratinocytes, Such feeder effects of the keratinocytes could be replaced to some extent with conditioned medium of the cells. Serial cultivation of papilla cells was established by maintaining them in the conditioned medium in which they were subcultured for more than 90 passages with an approximate population doubling time of 30 h, a value similar to that of rat dermal fibroblasts. During the subculture, they showed morphologic characteristics and phenotypic expressions of original papilla cells. Even after at least 70 passages, papilla cells sustained the innate hair follicle inductive ability at a level comparable with that of intact dermal papillae. The established cell lines did not show tumorigenicity when they were subcutaneously implanted into nude mice. The culture method developed in this study should facillitate the search for a biochemical entity of dermal papilla cells.