An improved organ culture for regeneration of pure autologous keratinocytes from small split-thickness skin specimens.
BACKGROUND
Failure of autologous keratinocyte culture from small split-thickness skin specimens or contamination of the keratinocyte culture by melanocytes represents practical problems in basic medical research and clinical studies.


OBJECTIVE
To establish a simple and reliable method of harvesting pure autologous keratinocytes from a small split-thickness skin specimen.


METHODS
Split-thickness (0.3 mm) skin explants (1 x 2 mm) were firstly cultured in DMEM containing 10% FCS till formation of keratinocyte strips, then cultured in serum-free keratinocyte growth medium or cocultured with lethally irradiated 3T3 fibroblasts (J2) in a mixture of DMEM and Ham's F12 (DF) medium.


RESULTS
Pure autologous keratinocyte culture is easily and reliably established by this organ culture technique.


CONCLUSIONS
Culturing of skin explants in serum-free keratinocyte growth medium or coculturing of the skin explants with lethally irradiated 3T3 cells in DF medium is proved to be a useful, simple and reliable method of harvesting pure autologous keratinocytes from a small split-thickness skin biopsy.
Liu
Jin Yu
JY
Department of Dermatology, University Hospital of Zurich, Zurich, Switzerland.
Burg
Gunter
G
D016475:3T3 Cells / Q000166:cytology; D000818:Animals; D001706:Biopsy / Q000379:methods; D003470:Culture Media; D006801:Humans; D015603:Keratinocytes / Q000166:cytology*; D007871:Leg Ulcer / Q000473:pathology; D051379:Mice; D009924:Organ Culture Techniques / Q000379:methods*; D012038:Regeneration; D012867:Skin / Q000166:cytology / Q000473:pathology; D012879:Skin Physiological Phenomena
D003470:Culture Media