Primary keratinocyte cell culture on EpiGen membranes for autologous skin grafts in paediatric burn patients

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South African Journal of Science

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University of Cape Town

Cultured epithelial autografts have been shown to be an effective permanent skin replacement for major burn injuries and have proved life-saving when insufficient donor skin has been available. Several membrane systems have been developed that facilitate the transfer of cultured cells on to the recipient. The aim of the study reported here was to test the effectiveness of EpiGen, a synthetic polymer membrane, as a cell culture support matrix for the transplantation of cultured autografts. Skin biopsies were obtained from 22 paediatric burn patients with an affected total body surface area of between 7% and 80%. Basal keratinocytes were harvested from the dermal/epidermal junction and cultured in a collagen 1-coated flask in modified Green’s medium. After two passages, isolated keratinocytes were grown on EpiGen membranes until semiconfluent. Wound beds were excised and covered with widely (1:3) meshed split skin grafts. Membranes were grafted with the basal cell layer directed against the wound bed. Unseeded membranes were applied and served as controls. Wounds were dressed and closed appropriately. Grafts were regularly inspected for ‘take’ and the membranes were removed 10 days after application. Seven patients had to be excluded from the study. Cell culture results of the remaining 15 patients showed excellent cell growth and expansion on EpiGen membranes within a mean culture time of 2.6 days post membrane seeding. The membranes facilitated easy transfer of cultures onto the recipient. A mean keratinocyte graft ‘take’ of 95% and a mean control graft ‘take’ of 90% were recorded at the time of membrane removal. The mean level of clinically evident re-epithelialization on keratinocyte grafted areas in recipients was 87% as opposed to 60% in the unseeded control areas.