SIGNALING PATHWAYS ACTIVATED DURING HEPATIC DIFFERENTIATION AND PROLIFERATION OF AMNIOTIC EPITHELIAL CELLS

RODRIGO RIEDEL; ANTONIO PÉREZ-PÉREZ; MARIANA JAIME; ORNELLA PAROLINI; ROBERTO CASALE; JOSÉ LUIS DUEÑAS; VICTOR SÁNCHEZ MARGALET; CECILIA VARONE; JULIETA MAYMÓ

Mar del Plata

Congreso; LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; 2019

SAIC

Abstract/Resumen: The placenta and fetal membranes haverecently been proposed as an important stem cells source forregenerative medicine. Amniotic epithelial cells (hAECs) can beisolated from the amnion of the human placenta at term. Theyexpress embryonic stem cells markers and they are pluripotent.These characteristics would make hAECs ideal candidates forregenerative medicine. Hepatic failure is one of the major causesof morbidity and mortality worldwide and the availabletreatments have several obstacles. Stem cells have beenspotlighted as alternative sources of hepatocytes because of theirpotential for hepatogenic differentiation. The adequate regulationof the signalling pathways activated during a differentiationprocess is the key for the success of such process. The aim ofthis work was to study some of the main pathways activated inhAECs during their hepatic differentiation process. Hepaticdifferentiation (HD) was assayed by specific HD medium (EGF +dexamethasone). Immunofluorescence (IF), Western blot (WB),qRT-PCR, PAS staining and MTT assays were performed. We havefound that HD significantly induced an increment in Wnt-1 and B-catenin expression in hAECs, measured by qRT-PCR, WB and IF.Treatment of hAECs with XAV939 (a B-catenin inhibitor) causedthe inhibition of HD, as albumin expression and glycogensynthesis were reduced. In addition, we determined that B-catenin pathway inhibition diminished Ki-67 expression and cellviability, during HD. We have also observed that HD promotesphosphorylation of PI3K and Akt, as determined by WB and IF.We observed a significant increment in nuclear localization of P-Akt during hAECs HD. These results suggest that the activationof the B-catenin and PI3K pathways may be responsible for asuccessful hepatic differentiation and proliferation of hAECs.Understanding the molecular mechanisms regulating hepatocytedifferentiation will significantly facilitate the development of stemcell-based therapy to treat liver diseases.