Establishment of Culture Technique of Keratinocytes from Plucked Human Hair Follicles

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  • 抜去毛包を試料とするヒトケラチノサイト培養系の確立

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Human epidermal keratinocyte strains are usually established from skin biopsies. Hair follicles may serve as an alternative source for keratinocytes, but the exact rate at which keratinocyte strains established using hair follicles had not been reported so far. Here we report an efficient and simple method to establish human keratinocyte strains using plucked hair follicles. Hair follicles with outer root sheath were obtained from adult scalps, placed in culture wells containing defined keratinocyte-serum free medium. The primary outgrowth was observed from 20 to 100% of follicles from every donor without regard to the sex or age. After two weeks of incubation, primary outgrowth from each follicle was trypsinized, transferred into a culture dish. Subcultures could be done for eighth to ninth passages to obtain 1.4×10^<11> to 7.3×10^<12> cells per follicle. The population doubling time was 28.4 h and the number of population doublings after the second passage was 26.2 on an average. The expression of the CD34 gene, one of the stem cell marker genes, was positive until the fourth passage. These results indicate that our method to establish keratinocyte strains from plucked hair follicles is efficient and applicable to somatic stem cell therapy.

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