Desarrollo, análisis y optimización de modelos celulares hepáticos para estudios de fármaco-toxicología y terapia celular

Resum

Given the importance of the liver in the metabolism and maintenance of the homeostasis of the organism, many studies have been conducted in the area of toxicology and, more recently, in hepatic cellular therapy. However, the main drawback is the limited availability of viable and functional hepatocytes due to the scarcity of liver tissue. The purpose of this work was based on the development and characterization of hepatic cellular models to become an alternative to hepatocytes in toxicology studies and cellular therapy. To this end, three main objectives have been investigated: 1) to adopt a procedure of hepatocyte isolation from discarded organs for transplantation which determines the optimal conditions for the isolation and culture of hepatocytes, 2) to characterize the cells from the hepatoblastome HepG2, and 3) to develop a hepatogenic differentiation protocol to induce the hepatic differentiation in adipose-derived stem cells (ADSC). In particular, the hepatogenic differentiation of stem cells opens a wide range of possibilities to facilitate the establishment of an adult differentiated cellular model useful for pharmaco-toxicological studies and for hepatic cellular therapy. The use of adult stem cells may allow the establishment of an adult cellular model with properties that others cellular models, like HepG2, do not show. However, it is necessary to optimise the isolation and cryopreservation procedures, as well as the differentiation protocols from adult stem cells and try to acquire a wide knowledge of the cellular and molecular mechanisms that control the transdifferentiation to hepatocytes.