Disappearance of Langerhans cells and melanocytes after cryopreservation of skin

Cryopreserved skin allografts have been extensively reported to remain viable for longer periods after grafting, both in the laboratory and in the clinic, than skin stored by other methods. We investigated the immunocytochemical and electron microscopic properties of samples of cryopreserved human skin (− 196°C) for comparison with fresh samples. In an immunocytochemical study of fresh skin, reagents S-100 and CDIa indicated numerous mesenchymal origin cells in the squamous cell layer, basal layer and dermis; 2B7 identified these cells in the basal layer and PC-10 identified them in the basal and squamous cell layers. In cryopreserved skin, however, few cells reacted to these reagents. An electron microscopic study of the cryopreserved skin showed Langerhans cells (LC); however, these had suffered degeneration, with partial defects of the cell membrane and vacuolation in the cytoplasm. We speculate these effects are responsible for the virtually complete abolition of LC membrane and cytoplasm markers. In summary, we detected few mesenchymal origin cells, melanocytes, Langerhans cells, or S-phase cells, in cryopreserved skin by immunocytochemical methods. Langerhans cells existed but had degenerated. These results indicate that cryopreservation at − 196°C causes degeneration of Langerhans cells, and that is the reason for the prolonged viability of cryopreserved allograft.