PROLIFERATION AND SURVIVAL DURING HEPATIC DIFFERENTIATION OF AMNIOTIC EPITHELIAL STEM CELLS

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

Buenos Aires

Congreso; Reunión Anual de las Sociedades de Biociencias 2017. LXI Reunión Anual de la Sociedad Argentina de Investigación Clínica (SAIC); 2017

Sociedades de Biosciencias

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, 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. After specific factors (HD) treatment, we observed a significant increase (2 ± 0.5 fold) in Cyclin D1 mRNA expression and a decrease (1.7 ± 0.3 fold) in p53 and (1.6 ± 0.4 fold) p21 expression, measured by qRT-PCR. This treatment also caused a down regulation (1.3 ± 0.5 fold) in p53 and p21 expression and an increment (1.9 ± 0.06 fold) 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 linked to cell growth and proliferation. Immunofluorescence and western blot analysis revealed the HD treatment significant increased (1.3 ± 0.1 fold) ERK 1/2 phosphorylation while CM diminished it (1.3 ± 0.04 fold). 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.