Cutaneous wound healing is a complex type of biological event involving proliferation, differentiation, reprograming, trans/de-differentiation, recruitment, migration, and apoptosis of a number of cells (keratinocytes, fibroblasts, endothelial cells, nerve cells and stem cells etc) to regenerate a multi-layered tissue that is damaged by either internal or external factors. Cell therapy has been proposed as a promising approach to be applied in clinics to have rapid and efficient repairing of wounds. A number of challenges like escape of cells, graft rejection etc are being faced for cell therapy to be a state-of-the-art approach for our daily routine clinical practice. Transdifferentiated stem cells (t-SCs) or keratinocyte-lineage committed stem cells (t-SCs) may show promising response in post-transplantation challenges such as cell escape as these cells have their lineage committed to engraft and develop the epidermis. Increased cell survival and less escape of transplanted cells is the current area of interest in regenerative medicine. Methods: Adipose-derived mesenchymal stem cells (ASCs) were cultured for transdifferentiation using three strategies as, 1) ASCs were cultured in keratinocytes conditioned medium (KCM) derived from the culture of rat keratinocytes cultured in keratinocytes growth medium (KGM), 2) co-culturing of ASCs with keratinocytes, and 3) ASCs were cultured in KGM supplemented with differentiation induction factors. ASCs were analysed for transdifferentiation after 7, 14 and 21 days of their culture for keratinocyte specific markers using RT-PCR. Differentiation assay, colony forming assay, and cell proliferation assay were performed to analyse differentiation, colonogenicity and proliferation potential of ASCs after 21 days of culturing. Results and Conclusion: Transdifferentiation of ASCs towards Keratinocyte-like stem cells was observed after 14 days as keratinocyte specific markers (K14, K18, K19) were expressed when ASCs were cultured in KCM, as co-culture with keratinocytes and cultured in induction medium for 7, 14 and 21 days. Transdifferentiated ASCs (t-ASCs) were found capable of differentiation, proliferation and clonogenic after 21 days of their culturing in KCM. It has been proposed that these t-SCs will be transplanted on rat models for cutaneous wound healing studies as it has been proposed that t-SCs toward keratinocyte-lineage are efficient in wound healing as compared to freshly derived ASCs or keratinocytes. Further investigations may prove the application of t-ASCs as the state-of-art approach to treat cutaneous wounds in preclinical and clinical studies.

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