title: Phenotypical characterization of 6-21-week gestational age human dermis and epidermal cell isolation methods for in vitro studies on epidermal progenitors.
abstract: Cell banked epidermal skin progenitor cells have the potential to provide an "off-the-freezer" product. Such cells may provide a skin donor area-independent cell-spray grafting therapy for the treatment of burns. We first characterized fetal skin samples of gestational ages ranging from 6 to 21 weeks. As the results suggest that the phenotypic differentiation occurs after 10 weeks, which may complicate follow-up in vitro studies, we developed and compared different cell isolation techniques for human fetal skin-derived epithelial cells from tissue ages 6 to 9 weeks. We initially screened seven methods of characterization, concluding that two methods warranted further investigation: incubating the epidermal tissue in Petri-dishes with culture medium for spontaneous cell outgrowth, and wiping the epidermal tissue onto a dry Petri-dish culture surface followed by adding culture medium. Non-controllable culture contamination with dermal cells was the reason for excluding the other five methods. The results suggest that epidermal cells can be isolated from tissue exhibiting a single homogeneous layer of CK15(+) basal keratinocytes up to week 9. At later gestational ages, the ongoing skin differentiation results in a multi-layer basal structure and progenitors associated with the hair bulb would have to be considered. Spraying the resulting cells with a clinical spray device was successfully demonstrated in an in vitro model.
  
  
  CONCLUSIONS
  Gestational age 6-9 weeks epidermal human fetal skin cells from the basal layer can be reproducibly isolated and transferred into culture for studies on the development of skin cell transplantation therapies.
authors: AG Experimental Surgery, BCRT, Charité Campus Virchow Clinic, University Medicine, Berlin, Germany. christa.johnen@charite.de
authors: Christa
authors: C
authors: Johnen
authors: Cinzia
authors: C
authors: Chinnici
authors: Fabio
authors: F
authors: Triolo
authors: Jörn
authors: J
authors: Plettig
authors: Kirsten
authors: K
authors: Bräutigam
authors: Giandomenico
authors: G
authors: Amico
authors: Matthew
authors: M
authors: Young
authors: Patrick
authors: P
authors: Over
authors: Roger
authors: R
authors: Esteban-Vives
authors: Eva
authors: E
authors: Schmelzer
authors: Pier-Giulio
authors: PG
authors: Conaldi
authors: Morris
authors: M
authors: Turner
authors: Robert
authors: R
authors: Thompson
authors: Katrin
authors: K
authors: Zeilinger
authors: Peter
authors: P
authors: Rubin
authors: Giovanni
authors: G
authors: Vizzini
authors: Bruno
authors: B
authors: Gridelli
authors: Jörg C
authors: JC
authors: Gerlach
mesh_terms: D002056:Burns / Q000601:surgery*; D018929:Cell Culture Techniques / Q000379:methods*; D017690:Cell Transplantation / Q000379:methods; D020405:Dermis / Q000166:cytology* / Q000196:embryology; D005865:Gestational Age; D006801:Humans; D016038:Skin Transplantation / Q000379:methods*; D013234:Stem Cells / Q000166:cytology*