Assessment of organ culture for the conservation of human skin allografts.
OBJECTIVE
Human skin allografts are used in the treatment of severe burns and their preservation is therefore critical for optimal clinical benefit. Current preservation methods, such as 4 degrees C storage or cryopreservation, cannot prevent the decrease of tissue viability. The aim of this study was to assess viability and function of skin allografts in a new skin organ culture model, allowing conservation parameters as close as possible to physiological conditions: 32 degrees C, air-liquid interface and physiological skin tension.


METHODS
Twelve skin samples, harvested from 6 living surgical donors, were conserved 35 days in two conditions: conservation at 4 degrees C and organ culture. Viability and function of skin samples were investigated at Day 0, 7, 14, 21, 28 and 35 using cell culture methods (trypan blue exclusion, Colony Forming Efficiency and Growth Rate), histopathological and histoenzymological studies (Ki67 immunostaining).


RESULTS
In the two conditions, fibroblast and keratinocyte viability was progressively affected by storage, with a significant decrease observed after 35 days. No statistical difference could be observed between the two conditions. The two methods were also comparable regarding alterations of fibroblast and keratinocyte culture parameters, which were respectively significantly reduced at Day 7 and 21, compared to fresh skin. By contrast, histopathological and histoenzymological studies revealed a better preservation of skin architecture and proliferative potential at 4 degrees C, as compared to organ culture.


CONCLUSIONS
These results indicate that skin organ culture does not provide significant advantages for skin allograft preservation. However, its potential use as an experimental model to study skin physiology and wound healing should be further evaluated.
Hautier
A
A
Service de Chirurgie Plastique, Centre des Grands Brûlés, Hôpital de La Conception, 147 boulevard Baille, Marseille, France. aurelie.hautier@ap-hm.fr
Sabatier
F
F
Stellmann
P
P
Andrac
L
L
Nouaille De Gorce
Y
Y
Dignat-George
F
F
Magalon
G
G
D000328:Adult; D049109:Cell Proliferation; D002470:Cell Survival / Q000502:physiology; D005260:Female; D005347:Fibroblasts / Q000473:pathology / Q000502:physiology; D006801:Humans; D015603:Keratinocytes / Q000473:pathology / Q000502:physiology; D008875:Middle Aged; D009924:Organ Culture Techniques / Q000379:methods*; D009926:Organ Preservation / Q000379:methods*; D012867:Skin; D016038:Skin Transplantation / Q000379:methods*; D013696:Temperature; D014184:Transplantation, Homologous / Q000379:methods