Human amniotic epithelial cells: Evaluation of survival during their hepatic differentiation

ANTONIO PÉREZ-PÉREZ; BERNARDO MASKIN; CECILIA LAURA VARONE; MALENA SCHANTON; ORNELLA PAROLINI; JULIETA LORENA MAYMÓ; RODRIGO RIEDEL; MARIANA JAIME; VICTOR SÁNCHEZ MARGALET

Manchester

Congreso; IFPA 2017; 2017

International Federation of Placenta Associations

The placenta and fetal membranes have recently been proposed as an important stem cells source for regenerative medicine. Amniotic epithelial cells isolated from human amnion (hAECs) offer considerable advantages that make them stand out between other stem cells. In addition to their virtually unlimited potential supply of, the easy access to such tissues, and the minimal ethical and legal barriers associated with their collection, they express embryonic stem cells markers and have the ability to differentiate toward all three germ layers. Moreover, they are not tumorigenic and have immunosuppressive properties. These characteristics would make hAECs ideal candidates for tissue engineering and application in regenerative medicine. Hepatic failure is one of the major causes of morbidity and mortality worldwide. Recently, stem cells have been spotlighted as alternative source of hepatocytes because their specific potential for differentiation. The aim of this work was to study the proliferation and survival of hAECs, during hepatic differentiation. Hepatic differentiation was assayed by specific factors (EGF + dexamethasone) or by HepG2 conditioned medium (CM). We have analyzed the expression of some key cell cycle proteins (Cyclin D1, p53 and p21). After specific factors treatment, we observed a significant increase in cyclin D1 mRNA expression and a decrease in p53 and p21 expression, measured by qRT-PCR. This treatment also caused a down regulation in p53 and p21 expression and an increment in cyclin D1, measured by western-blot. The opposite effects were observed with CM treatment. Finally, we have evaluated the MAPK signaling pathway activation, which is intimately linked to cell growth and proliferation. Immunofluorescence and western blot analysis revealed hepatic differentiation with specific factors significant increased ERK 1/2 phosphorylation while CM diminished it. Our results suggest hepatic differentiation with specific factors promotes the proliferation and survival of hAECs, improving their quality and quantity for an eventual future transplant.