Preservations in anhydrous sodium chloride reduces keratinocytes alloantigenicty. (P2223)

Abstract

Abstract A large amount of human keratinocytes (KCs) is needed to cover burned skin areas. There is a need to develop an efficient method of long term preservation of these cells. We checked whether KCs isolated from skin preserved in anhydrous sodium chloride survive and can be successfully transplanted. Fragments of normal human skin were preserved in sodium chloride. After 7 or 30 days preservation they were rehydrated, the epidermis was separated and KCs were isolated. Subsequently, the cells were cultured for 7 or 21 days. Viability of cultured KCs was tested by the Viability/Cytotoxicity Test. KCs were transplanted to the dorsum of SCID mice. Histopathological evaluation revealed that the morphology of KCs was totally preserved. KCs from all epidermal layers could be identified. All grafts were taken by the recipients. Three weeks after transplantation, keratinocytes synthesized keratins 10 and 16, and expressed antigens specific for stem and transient amplifying cells, as well as markers of proliferation. Cells isolated from transplants and cultured in vitro remained viable and produced various enzymes. All graft cells remained HLA I+. Taken together, keratinocytes isolated from human skin and preserved in anhydric sodium chloride present very low alloantigenicity can be successfully transplanted to SCID mice.