title: A new system for cultivation of human keratinocytes on acellular dermal matrix substitute with the use of human fibroblast feeder layer.
abstract: To improve the proliferative potential of human keratinocytes (HK) cultured on acellular dermal matrix (ADM), HK and mitomycin C-treated human fibroblasts (MMC-HF) were seeded onto ADM to form four types of composite skin: type I, HK were seeded onto the epidermal side of ADM; type II, both HK and MMC-HF were seeded onto the epidermal side; type III, MMC-HF were preseeded onto the dermal side of ADM, and then HK were seeded onto the epidermal side, and type IV, where MMC-HF were preseeded onto both sides, and then HK were seeded onto the epidermal side. Compared with type I and III, the proliferative potential of HK of type II and IV was significantly higher on day 3, 5, 7 and 9 in vitro. In type I and III, HK grew into one layer on day 7-9, while in type II and IV keratinocytes grew into a confluent monolayer by day 4-6. The adherence to ADM of HK in types II and IV was stronger than that in type I and III. The take rate of type II and IV composite skin was also significantly higher. In conclusion, when MMC-HF and HK were cocultured on the epidermal side of ADM, MMC-HF could serve as excellent feeder cells.
authors.affiliation: Burns Center, Changhai Hospital, Shanghai, PR China.
authors.forename: S
authors.initials: S
authors.lastname: Xiao
authors.forename: S
authors.initials: S
authors.lastname: Zhu
authors.forename: B
authors.initials: B
authors.lastname: Ma
authors.forename: Z-F
authors.initials: ZF
authors.lastname: Xia
authors.forename: J
authors.initials: J
authors.lastname: Yang
authors.forename: G
authors.initials: G
authors.lastname: Wang
mesh_terms: D000818:Animals; D001672:Biocompatible Materials; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D018920:Coculture Techniques; D024141:Collagen Type IV / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D006085:Graft Survival / Q000502:physiology; D006801:Humans; D007231:Infant, Newborn; D015603:Keratinocytes / Q000166:cytology* / Q000502:physiology; D007797:Laminin / Q000378:metabolism; D008297:Male; D051379:Mice; D008819:Mice, Nude; D016685:Mitomycin / Q000494:pharmacology; D016038:Skin Transplantation / Q000379:methods*; D017768:Skin, Artificial; D054457:Tissue Scaffolds
chemical_list: D001672:Biocompatible Materials; D024141:Collagen Type IV; D007797:Laminin; D016685:Mitomycin