Viability of cryopreserved human skin allografts: effects of transport media and cryoprotectant
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Viability of cryopreserved human skin allografts: effects of transport media and cryoprotectant Sonia Gaucher • Caroline Elie • Olivier Ve´rola Mohamed Jarraya
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Received: 1 September 2010 / Accepted: 13 January 2011 / Published online: 9 February 2011 Ó Springer Science+Business Media B.V. 2011
Abstract Human skin allografts can be preserved by different methods. In our clinical practice, human skin allografts are harvested on multi-organ and tissue donors, transferred at ?4°C in Ringer Lactate, cryopreserved with 15% Glycerol and held in the vapor phase of a liquid nitrogen freezer until delivery to the burn center. The aim of this experimental study was to evaluate the impact of transport medium and cryoprotectant on the viability of human skin allografts. For this purpose, we compared skin samples harvested from 19 multi-organ and tissue donors with S. Gaucher C. Elie Faculte´ de Me´decine, Universite´ Paris Descartes, Paris, France S. Gaucher (&) Service des Bruˆle´s, AP-HP Hoˆpital Cochin, 75014 Paris, France e-mail: [email protected] C. Elie Service de Biostatistique, AP-HP Hoˆpital Necker Enfants Malades, 75014 Paris, France O. Ve´rola Faculte´ de Me´decine, Universite´ Paris 7, Paris, France O. Ve´rola Service d’Anatomie Pathologie, AP-HP Hoˆpital Saint Louis, 75010 Paris, France M. Jarraya Banque des Tissus Humains, AP-HP Hoˆpital Saint Louis, 75010 Paris, France
two different transport media and two different cryoprotectants. Viability was assessed by the MTT assay after harvesting at laboratory reception, during storage (at ?4°C) at day 2 and day 7, and after cryopreservation and thawing. Histopathological analysis was performed for each MTT assay. Results indicate that, when stored at ?4°C, skin retains more viability with RPMI, whereas Glycerol and DMSO are equivalent cryoprotectants regardless of the transport medium. In conclusion, our protocol could be improved by the utilization of RPMI as transport medium. Keywords Human skin allografts Skin viability Transport medium Storage Cryopreservation MTT test Tissue banking
Introduction Early excision has been shown to be a major advance in the care of severe burn patients (Ong et al. 2006). When burn injuries are both deep and extensive, autografts are not available and skin substitutes have to be utilized. In this situation, human skin allografts, harvested from living or deceased donors, are considered one of the best temporary biologic coverages. The reasons are they limit hydro-electrolytic losses and pain through the mechanical effect on the wounds they cover; they reduce infection risk; and they boost tissue granulation, facilitating all the necessary factors for
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healing (Herndon and Parks 1986; Kreis et al. 1992; See et al. 2001; Naoum et al. 2004). Besides their temporary nature, the main drawback is the risk of disease transmission (Clarke 1987). Additional drawbacks in France are poor availability due to lack of donors and lack of harvesting. Tissue banks differ in their techniques for preservation and stor
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