CHANGES IN HUMAN AMNIOTIC EPITHELIAL STEM CELLS APOPTOSIS DURING THEIR HEPATIC DIFFERENTIATION

RODRIGO RIEDEL; ANTONIO PÉREZ-PÉREZ; NATALY DE DIOS; LUCIANO PÉREZ; MARIANA JAIME; ROBERTO CASALE; VICTOR SÁNCHEZ MARGALET; CECILIA VARONE; JULIETA MAYMÓ

CABA

Congreso; ? LXVII Reunión Anual de la Sociedad Argentina de Investigación Clínica; 2022

Sociedad Argentina de Investigación Clínica

CHANGES IN HUMAN AMNIOTIC EPITHELIAL STEM CELLS APOPTOSISDURING THEIR HEPATIC DIFFERENTIATIONRodrigo Riedel1, Antonio Pérez-Pérez2, Nataly de Dios1, Luciano Pérez1, MarianaJaime3, Roberto Casale3, Víctor Sánchez-Margalet2, Cecilia Varone1, Julieta Maymó1.1Biological Chemistry, IQUIBICEN, CONICET-FCEN, UBA, Ciudad Autónoma deBuenos Aires, Argentina; 2Medical Biochemistry and Molecular Biology andImmunology, Sevilla University, Sevilla, Spain; 3Maternity, Posadas Hospital, BuenosAires, Argentina.Amniotic epithelial stem cells can be isolated from the human placenta at term. Theyexpress embryonic stem cells markers, and they are pluripotent. Moreover, they do notexpress telomerase, they are not tumorigenic, and they have immunosuppressiveproperties. These characteristics position hAECs as ideal candidates for regenerativemedicine. Hepatic failure is one of the major causes of morbidity and mortality worldwideand the available treatments have several obstacles. Recently, hAECs have beenspotlighted as an alternative source of hepatocytes because of their potential forhepatogenic differentiation. This work aimed to assess the changes in hAECs apoptosisduring hepatic differentiation. Previously, we have demonstrated that hAECs efficientlydifferentiate to hepatic-like cells, by applying a specific hepatic differentiation (HD)protocol. We found that HD medium enhances proliferative capacity of hAECs. In thisway, we showed that HD significantly increases PCNA expression, measured by WesternBlot. We have analyzed the expression of some key apoptosis proteins (Caspase-8,Caspase-3, PARP-1) by qRT-PCR and Western blot. We have also evaluated p53expression by immunofluorescence. We found a significant reduction in cleaved Caspase-3 and PARP-1 during hAECs HD. Moreover, Caspase-8 expression significantlydiminishes in control hAECs while HD treatment prevented this effect. Additionally, wehave showed a significant decrease in p53 nuclear localization during hAECs HD,evaluated by immunofluorescence. Finally, we determined that HD treatment decreasesthe apoptotic nucleus number, measured by DAPI staining. Our results suggest that ourhepatic differentiation method not only induces proliferation and survival of hAECs butalso reduces apoptosis rates, improving their quality and quantity for an eventual futuretransplant.