Questions under study: Composite skin containing autologous keratinocytes is a new approach to solving the problem of extensive skin defects. We propose a new strategy to construct the composite skin by mixing autologous and non-autologous keratinocytes, so that the time and the cost for constructing the composite skin could be reduced.
Methods: Human keratinocytes were mixed with Balb/c mouse keratinocytes at appropriate proportions, seeded onto the surface of porcine acellular dermal matrix (ADM) and cultured in vitro to reconstruct composite skin, which was then transplanted to a Balb/c mouse skin defect. Quality of the wound healing as well as the homing of the non-autologous keratinocytes were observed.
Results: Wounds healed well with the transplanted composite skin containing two different keratinocytes. The take rate of the grafts ranged from 78.3% to 81.5%. Histological observation showed that both epidermal and dermal structures of the regenerated skin were perfect and that the basal membrane was obvious with the laminin and collagen IV positively stained. In the early grafting phase, there are many non-autologous keratinocytes in the new epidermis. With the lapse of time, non-autologous keratinocytes decreased gradually and were eventually replaced by the autologous keratinocytes.
Conclusion: A composite skin was reconstructed by mixing two different keratinocytes at a certain proportion and then co-culturing them with dermal scaffold, which can be used for repairing full thickness skin defect wounds. This new strategy could save the source of autologous skin effectively and shorten in vitro culture time of composite skin.