Pre-Differentiated Skin-Like Cells can Alleviate In Vitro Burn Injury

Abstract

Considering importance of stem cell based therapies for wound healing, we have previously established protocols for successful differentiation of stem cells (isolated from placenta) into skin-like cells (keratinocytes and fibroblasts). In the current study we aim to evaluate the effect of pre-differentiated skin-like-cells on wound healing potential using an in vitro burn injury model. The amniotic epithelial cells (AECs) and umbilical cord mesenchymal stem cells (UCMSCs) obtained from amniotic membrane and umbilical cord tissue; respectively, were differentiated into skin-like cells (keratinocyte and fibroblasts respectively). In order to make an in vitro burn injury model, keratinocytes and fibroblasts were isolated from rats and insulted with high temperature (up to 50oC). The optimal temperature for burn injury models was determined using viability assay, cytotoxicity level, proliferation, and expression of pro-apoptotic. In order to assess effectiveness of pre-differentiated skin like cells, the keratinocyte- and fibroblasts-like cells were co-cultured with injured keratinocytes and injured fibroblasts, respectively. Results indicated that optimal temperature for the induction of heat injury for both keratinocytes and fibroblasts was 50oC. At this temperature both types of cells (keratinocytes and fibroblasts) showed modified morphology, drastic cellular injury, least viability, higher expression of pro-apoptotic markers and lower expression of proliferation genes. Co-culturing of pre-differentiated cells leads to an increase in viability and proliferation concurrent with decreased apoptosis. We have established successfully in vitro model of burn injury that might be helpful to evaluate the wound healing potential of cells. Further, predifferentiated skin-like cells are a potential source for the treatment of severe burn injuries.