For hair follicle regeneration, the bioengineered hair germs were ectopically engrafted into the subrenal capsules of 8-week-old C57BL/6 mice, as previously described
7,
13, or they were intracutaneously transplanted onto the back skin of 6-week-old Balb/c nu/nu mice. Shallow stab wounds, which were nearly parallel to the skin surface and ~0.4 mm vertical and 2.8 mm horizontal from the needle-stick point, were made in the back skin of nude mice with a 20-G Ophthalmic V-Lance (Alcon Japan, Tokyo, Japan). The bioengineered hair follicle germ, which contained a nylon guide, was arranged in the epithelial portion and above, and it was intradermally transplanted into the wounds. The nylon guide was held so that it protruded from the skin surface. The transplantation sites were then covered with surgical bandage tape (Nichiban, Tokyo, Japan). To examine the engraftment of the transplants and to study the hair cycles of the bioengineered hairs, all transplanted sites were observed using a SteREO Lumar V12 and AxioCam fluorescent stereoscopic microscope system (Carl Zeiss, Oberkochen, Germany). Observations were made every 2–3 days.
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