title: Cultured epidermal keratinocytes on a microspherical transport system are feasible to reconstitute the epidermis in full-thickness wounds.
abstract: Research efforts to modify cultured autologous skin transplants for large full-thickness burn wounds and in chronic ulcers have shifted from multilayered differentiated grafts ("sheet" grafts) toward smaller units of basal undifferentiated single cell suspensions in a transport medium and subconfluently covered static carriers. It has been shown that wounds transplanted with single cell suspensions reconstitute the epidermis. However, this technique requires the detachment of the keratinocytes from the culture flasks by enzymatic digestion-digestion that might alter the anchoring proteins of the cells. A new approach might be to circumvent the enzymatic digestion to harvest the keratinocytes. This study reports a technique to culture epidermal cells on spherical microcarriers as a suspension culture and transport vehicle. The spherical microcarrier consists of a 100-microm-diameter collagen-coated dextran carrier (Cytodex 3 Pharmacia) and has been used previously for enzyme production commercially. With this new approach, we seeded the human keratinocytes in a spinner-like system onto microspheres and transplanted these micrografts onto full-thickness wounds on the back of nude mice. After 14 days, we showed a reconstituted epithelium that was multilayered and keratinized compared to control wounds. We believe that this is the first step of a new approach to increase the cell yield for seeding without altering the anchoring proteins by enzymatic steps, leading to a superior transplantation method for keratinocytes.
authors.affiliation: Department of Plastic and Hand Surgery, University of Freiburg Medical Center, Freiburg, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Voigt
authors.forename: M
authors.initials: M
authors.lastname: Schauer
authors.forename: D J
authors.initials: DJ
authors.lastname: Schaefer
authors.forename: C
authors.initials: C
authors.lastname: Andree
authors.forename: R
authors.initials: R
authors.lastname: Horch
authors.forename: G B
authors.initials: GB
authors.lastname: Stark
mesh_terms: D000328:Adult; D000818:Animals; D001415:Back; D018929:Cell Culture Techniques / Q000295:instrumentation*; D002455:Cell Division; D017690:Cell Transplantation / Q000295:instrumentation*; D002478:Cells, Cultured / Q000637:transplantation; D003094:Collagen; D015925:Cryopreservation; D016895:Culture Media, Serum-Free; D003911:Dextrans; D004817:Epidermis / Q000293:injuries* / Q000473:pathology; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000637:transplantation*; D051379:Mice; D008819:Mice, Nude; D008863:Microspheres; D014183:Transplantation, Heterologous; D014945:Wound Healing
chemical_list: D016895:Culture Media, Serum-Free; D003911:Dextrans; D003094:Collagen