title: Skin cell isolation and expansion for cell transplantation is limited in patients using tobacco, alcohol, or are exhibiting diabetes mellitus.
abstract: The aim of this exploratory study was to investigate the isolation and expansion of keratinocytes and fibroblasts from donors with certain medical histories. Biopsies were taken from donors (N=32) falling into one or more of the following categories: a history of heavy smoking and/or alcohol abuse, drug abuse, diabetes mellitus or steroid treatment. Cells from donors who did not fall into any of the above-mentioned categories were used as controls. Proliferation and growth behaviour of cells were analyzed by measurement of passage duration, absorbance (MTT-assay) and light microscopy. Donors with a specific medical history required larger biopsy areas than the control group for isolating a sufficient number of fibroblasts and keratinocytes. Times to confluence were significantly prolonged and absorbances (MTT) were significantly reduced in several donor groups when compared to control cultures. Biopsies from donors with steroid treatment, drug abuse and combined nicotine and alcohol abuse could not be established beyond passage 0 degrees or 1 degree, respectively. We conclude that isolation and expansion of skin cells from donors with certain medical histories may require larger biopsies, prolonged expansion times or may even result in failure. These findings may therefore be of clinical importance in the field of autologous skin cell transplantation.
authors.affiliation: Charité, Campus Virchow-Clinic, Department of Surgery, Universitäts Medizin Berlin, Germany.
authors.forename: Christa
authors.initials: C
authors.lastname: Johnen
authors.forename: Bernd
authors.initials: B
authors.lastname: Hartmann
authors.forename: Ingo
authors.initials: I
authors.lastname: Steffen
authors.forename: Kirsten
authors.initials: K
authors.lastname: Bräutigam
authors.forename: Tom
authors.initials: T
authors.lastname: Witascheck
authors.forename: Nidal
authors.initials: N
authors.lastname: Toman
authors.forename: Markus V
authors.initials: MV
authors.lastname: Küntscher
authors.forename: Jörg C
authors.initials: JC
authors.lastname: Gerlach
mesh_terms: D000328:Adult; D000368:Aged; D000437:Alcoholism / Q000150:complications / Q000473:pathology*; D001707:Biopsy, Needle / Q000379:methods; D002056:Burns / Q000601:surgery; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D000075202:Contraindications; D048909:Diabetes Complications / Q000150:complications / Q000473:pathology*; D005260:Female; D005347:Fibroblasts / Q000473:pathology* / Q000637:transplantation; D006801:Humans; D015603:Keratinocytes / Q000473:pathology* / Q000637:transplantation; D008297:Male; D008875:Middle Aged; D016038:Skin Transplantation / Q000473:pathology; D012907:Smoking / Q000009:adverse effects / Q000473:pathology*