title: A Two-Stepped Culture Method for Efficient Production of Trichogenic Keratinocytes.
abstract: Successful hair follicle (HF) neogenesis in adult life depends on the existence of both capable dermal cells and competent epidermal keratinocytes that recapitulate embryonic organogenesis through epithelial-mesenchymal interaction. In tissue engineering, the maintenance of trichogenic potential of adult epidermal cells, while expanding them remains a challenging issue. We found that although HF outer root sheath keratinocytes could be expanded for more than 100 passages as clonogenic cells without losing the proliferative potential with a 3T3J2 fibroblast feeder layer, these keratinocytes were unable to form new HFs when combined with inductive HF dermal papilla (DP) cells. However, when these high-passage keratinocytes were cocultured with HF DP cells for 4 days in vitro, they regained the trichogenic ability to form new HFs after transplantation. We found that the short-term coculture with DP cells enhanced both Wnt/β-catenin signaling, a signaling cascade key to HF development, and upregulated the expression of HF-specific genes, including K6, K16, K17, and K75, in keratinocytes, indicating that these cells were poised toward a HF fate. Hence, efficient production of trichogenic keratinocytes can be obtained by a two-stepped procedure with initial cell expansion with a 3T3J2 fibroblast feeder followed by short-term coculture with DP cells.
authors.affiliation: 1 Institute of Biomedical Engineering, National Taiwan University , Taipei, Taiwan .|2 Department of Dermatology, National Taiwan University Hospital and College of Medicine , Taipei, Taiwan .
authors.forename: Chih-Chieh
authors.initials: CC
authors.lastname: Chan
authors.affiliation: 1 Institute of Biomedical Engineering, National Taiwan University , Taipei, Taiwan .
authors.forename: Sabrina Mai-Yi
authors.initials: SM
authors.lastname: Fan
authors.affiliation: 1 Institute of Biomedical Engineering, National Taiwan University , Taipei, Taiwan .
authors.forename: Wei-Hung
authors.initials: WH
authors.lastname: Wang
authors.affiliation: 2 Department of Dermatology, National Taiwan University Hospital and College of Medicine , Taipei, Taiwan .
authors.forename: Yi-Fen
authors.initials: YF
authors.lastname: Mu
authors.affiliation: 1 Institute of Biomedical Engineering, National Taiwan University , Taipei, Taiwan .|2 Department of Dermatology, National Taiwan University Hospital and College of Medicine , Taipei, Taiwan .
authors.forename: Sung-Jan
authors.initials: SJ
authors.lastname: Lin
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000096:biosynthesis*; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D061252:Feeder Cells / Q000166:cytology / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation; D018859:Hair Follicle / Q000166:cytology / Q000378:metabolism*; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism* / Q000637:transplantation; D051381:Rats; D017208:Rats, Wistar
chemical_list: D000943:Antigens, Differentiation