Effective Strategies for Modulation of Amniotic Epithelial Stem Cells' Epithelial-Mesenchymal Transition in vitro

Author: Merabi Sepashvili
Keywords: Regenerative medicine, Stem cells, Amniotic epithelial cells, Epithelial-mesenchymal transition, Progesterone, TGF-β
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Regenerative medicine holds great promise for the development of innovative therapies aimed at organ regeneration and tissue repair. Stem cells, with their ability to differentiate into various cell types, are a key focus of regenerative medicine research. Amniotic epithelial cells (AECs), derived from the amniotic fluid and membrane, have emerged as a promising candidate due to their unique properties. These cells express embryonic and pluripotency markers, exhibit low immunogenicity, and possess strong immunomodulatory activity. They can be obtained without ethical and legal concerns making them an appealing option for therapeutic applications. During in vitro expansion, AECs experience spontaneous epithelial-mesenchymal transition (EMT) which involves various molecular and cellular changes, including alterations in cell morphology, cytoskeletal reorganization, and changes in gene expression patterns. Understanding the molecular mechanisms and signaling pathways associated with EMT in AECs is essential for controlling their behavior and enhancing their therapeutic potential. This study aims to evaluate effective strategies for modulating EMT in AECs in vitro. The specific objectives of this study include the isolation and cultivation of ovine amniotic epithelial cells (oAECs), morphological analysis of oAECs, evaluation of epithelial and mesenchymal phenotype cell markers using immunofluorescent analysis, and determination of the optimal concentration of TGF-β for oAECs’ EMT stimulation in vitro. The present thesis provides further evidence for the capacity of TGF-β and progesterone (P4) to induce phenotypical alterations of oAECs in vitro. Furthermore, we demonstrate that TGF-β, similar to P4, exerts a dose-dependent effect on epithelial-mesenchymal transition of oAECs in vitro, highlighting its regulatory role in this process. Thus, through an interdisciplinary approach combining molecular biology, cell culture techniques, and advanced imaging methods, this research contributes to the better understanding of AECs and their potential in regenerative medicine.