The Creation of Human Epithelial Tissue Graft Derived From Human Blood Cord Mesenchymal Stem Cells and Human Amniotic Membrane to Be Applied in Emergency Care

Abstract

Due to its outstanding biological properties human amniotic membrane (hAM) has been a topic of medical research for many years in trying to treat, repair and regenerate difficult damages related to the epithelial surface, corneal epithelial, orthopedic, and infected body parts. The hAM’s showed also to possess crucial anti-inflammatory, anti-bacterial, and anti-viral properties and immunomodulating qualities. As a consequence, the hAM is one of the most used natural bio-materials in tissue engineering for the construction of bio-scaffolds. We used human hAM (hAM) as a raw material source to home umbilical cord mesenchymal stem cells (hUCB-MSCs) to manufacture collagen membranes that could fully meet the standards for making a skin scaffold, with suitable mechanical strength whilst limiting the risk graft rejection. Among the varieties of stem cells currently being researched MSCs are the most studied and applied due to their outstanding bio-qualities and regenerative features. We used International Society for Cells and Gene Therapy (ISCT) standards to identify and characterize the hUCB-MSCs obtained from a consent donor’s umbilical cord blood. In this study, we used hAM as a scaffold to culture domestic hUBC-MSCs to create human-like epithelial tissues aimed at repairing and regenerating the epithelial layer following trauma, deep damage from accidents, burns, and/or conditions post-operative. We used an inverted phase contrast microscope to observe the cultured growing cells on the amniotic specimens to evaluate morphological changes and epithelial stratification. Staining techniques such as H&E and Trichrome were used to evaluate the morphology and structure of the new epithelial construct, histochemical staining with p63 and ck 1/10 markers was instead used to evaluate the cellular characteristics, the transmission electron microscopy (TEM) was used to validate cell junctions. Ultimately, the results successfully showed the possibility of obtaining human-like epithelial tissue with multiple cell layers with morphology and characteristics similar to human epithelial-like tissue. The results of this study will constitute a potential alternative to autologous skin grafting in cases of heavy skin loss and deep lesions.