Waldemar L Olszewski1, Maria Moscicka, Dorota Zolich. 1. Department of Surgical Research and Transplantology, Medical Research Center, Polish Academy of Sciences, Warsaw, Poland. wlo@cmdik.pan.pl
Abstract
BACKGROUND: Human skin is needed to cover large areas of the body lost through burns, trauma, and extensive maxillofacial surgery. Contemporary methods of skin storage are limited by the period of preservation to a few days. Our previous findings showed that fixation and storage of human skin in anhydric sodium chloride at room temperature for weeks or months preserves its morphological and molecular structure. In this study, we examined whether skin grafts preserved in sodium chloride may be successfully transplanted. METHODS: Skin was harvested from lower limbs of patients during elective surgery, placed in containers with anhydric salt powder, and kept at 22 degrees C for 3 to 12 weeks. Desalination and rehydration took place before transplantation. Desalinated fragments were transplanted onto the dorsum of scid mice. RESULTS: All grafts were accepted by recipients. Three weeks after transplantation, keratinocytes synthesized keratins 10, 16, and 17 and expressed antigens specific for stem (p63) and transient (CD29) cells. Moreover, they proliferated vigorously, their basal layer cells incorporated bromdeoxyuridine and expressed proliferative cell nuclear antigen. Isolated from transplants and cultured in vitro, they remained viable and produced enzymes. Dermis retained its structure and expressed fibroblast-specific antigen. All graft cells remained human leukocyte antigen I. CONCLUSION: Human skin preserved in anhydric sodium chloride at room temperature for months can be successfully transplanted to scid mice. We propose the concept of "spore-like" keratinocyte stem cells to explain the long-term ex vivo survival of keratinocytes. The mechanism of survival of fibroblasts remains to be determined.
BACKGROUND:Human skin is needed to cover large areas of the body lost through burns, trauma, and extensive maxillofacial surgery. Contemporary methods of skin storage are limited by the period of preservation to a few days. Our previous findings showed that fixation and storage of human skin in anhydric sodium chloride at room temperature for weeks or months preserves its morphological and molecular structure. In this study, we examined whether skin grafts preserved in sodium chloride may be successfully transplanted. METHODS: Skin was harvested from lower limbs of patients during elective surgery, placed in containers with anhydric salt powder, and kept at 22 degrees C for 3 to 12 weeks. Desalination and rehydration took place before transplantation. Desalinated fragments were transplanted onto the dorsum of scid mice. RESULTS: All grafts were accepted by recipients. Three weeks after transplantation, keratinocytes synthesized keratins 10, 16, and 17 and expressed antigens specific for stem (p63) and transient (CD29) cells. Moreover, they proliferated vigorously, their basal layer cells incorporated bromdeoxyuridine and expressed proliferative cell nuclear antigen. Isolated from transplants and cultured in vitro, they remained viable and produced enzymes. Dermis retained its structure and expressed fibroblast-specific antigen. All graft cells remained human leukocyte antigen I. CONCLUSION:Human skin preserved in anhydric sodium chloride at room temperature for months can be successfully transplanted to scid mice. We propose the concept of "spore-like" keratinocyte stem cells to explain the long-term ex vivo survival of keratinocytes. The mechanism of survival of fibroblasts remains to be determined.