title: In vitro keratinocyte expansion for cell transplantation therapy is associated with differentiation and loss of basal layer derived progenitor population.
abstract: An alternative approach for traditional clinical mesh grafting in burn wound treatment is the use of expanded autologous keratinocytes in suspension or sheets that are cultured over 2-4 weeks in a remote service facility. While a wound reepithelialization has been described, the functional and aesthetic outcome is under debate. Cell isolation from split-skin donor tissue aims to preserve the valuable stem cell progenitors from the basal epidermal layer and to provide patients with a rapid wound reepithelialization and a satisfying outcome. While the presence of epidermal progenitors in the cell graft is thought to enable an improved epidermal surface post reepithelialization, we investigated a feasible clinical approach involving cultured versus noncultured epidermal cells comparing the α6int(high)/K15(high)/FSC(low)/SSC(low) and α6int(high)/K5(high)/FSC(low)/SSC(low) keratinocyte progenitor subpopulations before and after in vitro culture process. Our results show a significant increase of cell size during in vitro passaging and a decrease of progenitor markers linked to a gradual differentiation. A provision of the regenerative epidermal progenitors, isolated from the split-skin biopsy and applied directly onto the wound in an on-site setting of isolation and cell spray grafting in the operation room, could be of interest when choosing options for skin wound care with autologous cells.
authors.affiliation: Departments of Surgery and Bioengineering, McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA. Electronic address: roger.esteban@gmail.com.
authors.forename: Roger
authors.initials: R
authors.lastname: Esteban-Vives
authors.affiliation: Departments of Surgery and Bioengineering, McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Matt
authors.initials: M
authors.lastname: Young
authors.affiliation: Departments of Surgery and Bioengineering, McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Patrick
authors.initials: P
authors.lastname: Over
authors.affiliation: Departments of Surgery and Bioengineering, McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Eva
authors.initials: E
authors.lastname: Schmelzer
authors.affiliation: University of Pittsburgh Medical Center, UPMC Mercy Hospital Trauma Services and Burn Unit, Pittsburgh, PA, USA.
authors.forename: Alain
authors.initials: A
authors.lastname: Corcos
authors.affiliation: University of Pittsburgh Medical Center, UPMC Mercy Hospital Trauma Services and Burn Unit, Pittsburgh, PA, USA.
authors.forename: Jenny
authors.initials: J
authors.lastname: Ziembicki
authors.affiliation: Departments of Surgery and Bioengineering, McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Jörg
authors.initials: J
authors.lastname: Gerlach
mesh_terms: D000328:Adult; D002454:Cell Differentiation; D049109:Cell Proliferation; D048429:Cell Size; D017690:Cell Transplantation; D064987:Cell- and Tissue-Based Therapy; D002478:Cells, Cultured; D004817:Epidermis / Q000637:transplantation*; D006801:Humans; D015603:Keratinocytes / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology
keywords: Cell culture; Differentiation; Keratinocytes; Progenitor; Wound healing