Changes in cellular properties during hepatic differentiation of hAECs

MAYMÓ J; PEREZ PEREZ A; MAGATTI M; MASKIN B; PAROLINI O; SÁNCHEZ MARGALET V; VARONE C

Granada

Congreso; 3° IPLASS Meeting: Towars clinical applications of placental and endometrial stem cells; 2014

IPLASS

The placenta and fetal membranes have recently been proposed as an important stem cells source for regenerative medicine. Gestational cells offer considerable advantages over other stem cells such as bone marrow or embryo-derived cells. There is a virtually unlimited potential supply of, an easy access to such tissues, and minimal ethical and legal barriers are associated with their collection and use. Epithelial amniotic cells (hAECs) can be isolated from the amnion of the human placenta at term. They express embryonic stem cells markers and have the ability to differentiate toward all three germ layers. Moreover, their immunosuppressive properties might render allogenic application possible. 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. Although it is the best way to treat liver transplantation for acute and chronic hepatic failure patients, there are several obstacles. Recently, stem cells have been spotlighted as alternative sources of hepatocytes because their potential for hepatogenic differentiation. The aim of this work was to study the proliferation and apoptosis of hAECs, during early and late hepatic differentiation. We have also analyzed pluripotent genes (Sox-2, Oct-4, Nanog) and hepatic markers expression through this process. Hepatic differentiation was assayed by specific factors (EGF + dexamethasone) or by HepG2 conditioned medium (CM). We have found an increment of caspase-3 fragmentation, after 72 h of hAECs treatment, measured by western blot. We have also determined by 3H-thymidine incorporation, that EGF significant increased cell proliferation while CM diminished it. During hAECs differentiation we observed by qRT-PCR, a significant decrease in pluripotent genes expression and an increment in hepatocytes-related genes (alfa-fetoprotein, alfa1-AT, albumin, CYP7A1). Our results begin to unravel molecular and cellular process that take place during hepatic differentiation of hAECs.