title: Method for autologous single skin cell isolation for regenerative cell spray transplantation with non-cultured cells.
abstract: BACKGROUND
  There is a therapeutic gap for patients with deep partial thickness wounds (Grade IIb) of moderate size that were initially not treated with split- or mesh grafting to avoid overgrafting, but developed delayed wound healing around two weeks after injury--at which time grafting is typically not indicated anymore. Delayed wound healing is often associated with esthetically unsatisfactory results and sometimes functional problems. An innovative cell isolation method for cell spray transplantation at the point of care, which eliminates cell culture prior to treatment, was implemented for this population of burn patients in our center.
  
  
  METHODS
  Autologous skin cell spray transplantation was initiated by taking healthy skin. The dermal/epidermal layers were separated using enzymatic digestion with 40 min dispase application, followed by 15 min trypsin application for basal kerationcyte isolation, 7 min cell washing by centrifugation, followed by transferring the cells for spraying into Ringer lactate solution. The procedure was performed on site in a single session immediately following the biopsy. After sharp wound debridement, cells were immediately transplanted by deposition with a cell sprayer for even distribution of the cell suspension.
  
  
  CONCLUSIONS
  Eight patients were treated (mean age 30.3 years, mean burn total body surface area 14%, mean Abbreviated Burn Severity Index (5 points). The mean time to complete re-epithelialization was 12.6 days. All patients exhibited wound healing with improved esthetic and functional quality. Our initial experience for the use of non-cultured cells using a two-enzyme approach with cell washing suggests shortened time for wound closure, suggesting that the method may potentially avoid longer-term complications.
authors.affiliation: Department of Surgery, McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania 15203, USA. joerg.gerlach@cellnet.org
authors.forename: Jörg C
authors.initials: JC
authors.lastname: Gerlach
authors.forename: Christa
authors.initials: C
authors.lastname: Johnen
authors.forename: Christian
authors.initials: C
authors.lastname: Ottomann
authors.forename: Christian
authors.initials: C
authors.lastname: Ottoman
authors.forename: Kirsten
authors.initials: K
authors.lastname: Bräutigam
authors.forename: Jörn
authors.initials: J
authors.lastname: Plettig
authors.forename: Claudia
authors.initials: C
authors.lastname: Belfekroun
authors.forename: Sandra
authors.initials: S
authors.lastname: Münch
authors.forename: Bernd
authors.initials: B
authors.lastname: Hartmann
mesh_terms: D000293:Adolescent; D000328:Adult; D002056:Burns / Q000601:surgery*; D017690:Cell Transplantation / Q000379:methods*; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D008875:Middle Aged; D012867:Skin / Q000166:cytology*; D016038:Skin Transplantation / Q000379:methods*; D016896:Treatment Outcome; D014945:Wound Healing / Q000502:physiology*