Assessment of organ culture for the conservation of human skin allografts
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Assessment of organ culture for the conservation of human skin allografts A. Hautier Æ F. Sabatier Æ P. Stellmann Æ L. Andrac Æ Y. Nouaille De Gorce Æ F. Dignat-George Æ G. Magalon
Received: 24 September 2006 / Accepted: 28 March 2007 / Published online: 26 April 2007 Springer Science+Business Media B.V. 2007
Abstract 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 48C 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: 328C, air–liquid interface and physiological skin tension. Design Twelve skin samples, harvested from 6 living surgical donors, were conserved 35 days in two conditions: conservation at 48C and organ A. Hautier (&) G. Magalon Service de Chirurgie Plastique, Centre des Grands Bruˆle´s, Hoˆpital de La Conception, 147 boulevard Baille, Marseille, France e-mail: [email protected] F. Sabatier P. Stellmann F.Dignat-George Centre de The´rapie Cellulaire, Hoˆpital de La Conception, Marseille, France
L.Andrac Laboratoire d’Anatomie et de Cytologie Pathologique, Faculte´ de Me´decine Nord, Marseille, France Y. Nouaille De Gorce Banque de Tissus EFS Alpes-Me´diterrane´e, Marseille, France
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 48C, as compared to organ culture. Conclusion 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. Keywords Cell culture Conservation Histopathology Organ culture Physiological skin tension Skin allografts Tissue banking Abbreviations CFE Colony Forming Efficiency CFU Colony Forming Unit DMEM Dulbecco’s Modified Eagle Medium
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20
FBS MTT PBS PD
Cell Tissue Banking (2008) 9:19–29
Fetal Calf Serum Methyl Thiazolil Tetrazolium Phosphate Buffered Saline Population Doubling
strongly reduces after harvest, due to dermal elasticity.
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