Characterization of hair follicles induced in implanted, cultured rat keratinocyte sheets.
Cultured rat keratinocyte sheets form hair follicles in combination with rat vibrissa dermal papillae when they are transplanted subcutaneously in syngeneic rats and athymic mice. In the present study, the histologic details of these induced follicles were analyzed by preparing cultured sheets mixed with normal rat keratinocytes and green fluorescent protein (GFP)-transgenic rat keratinocytes. Histologic examination demonstrated that some induced follicles maintained their size and morphology for at least 18 weeks, whereas others decreased in size and others totally differentiated into cornified structures between 3 and 6 weeks. The percentage of the grafts with GFP-positive cells decreased during the same period. This finding suggests that some GFP-positive cells were transient-amplifying cells that turned into terminally differentiated cells and were lost during this period. Some large follicles and some small follicles maintained their hair-producing ability and the proliferative activity in their hair matrix for 18 weeks. In addition, one 6-week-old follicle contained label-retaining cells in the outer root sheath. Seven of 25 follicles induced from chimera epithelium contained both GFP-positive cells and GFP-negative cells. These results suggest that stem cells are present in the induced follicle and the induced follicle consists of polyclonally derived cells. The presence of early anagen-like large follicles at week 6 and 9 and a telogen-like small follicle at week 18 also suggests that hair-growth cycle phases proceeded in the induced follicles. In conclusion, the follicles induced in the cultured keratinocyte sheets maintained hair-producing ability and proliferative activity for at least 18 weeks. This and the presence of label-retaining cells suggest that there are stem cells in the induced follicles, which seem to have a hair-growth cycle.
Miyashita
H
H
Graduate School of Science and Technology, Chiba University, Chiba, Japan.
Hakamata
Y
Y
Kobayashi
E
E
Kobayashi
K
K
D000818:Animals; D030801:Animals, Genetically Modified; D001973:Bromodeoxyuridine / Q000494:pharmacology; D017690:Cell Transplantation; D002478:Cells, Cultured; D004396:Coloring Agents / Q000494:pharmacology; D004247:DNA / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018859:Hair Follicle / Q000378:metabolism*; D007150:Immunohistochemistry; D015603:Keratinocytes / Q000378:metabolism*; D051379:Mice; D008819:Mice, Nude; D051381:Rats; D011916:Rats, Inbred F344; D013234:Stem Cells; D013997:Time Factors
D004396:Coloring Agents; D049452:Green Fluorescent Proteins; D004247:DNA; D001973:Bromodeoxyuridine