Obtaining hepatocyte-like cells from dental pulp pluripotent-like stem cells

Abstract

The liver is the largest internal organ and an important regulator of homeostasis providing essential metabolic, exocrine and endocrine functions. Liver diseases result in high rates of morbidity and mortality and affect a growing number of people over the years. Obtaining mature hepatocytes from stem cells is a major goal in regenerative medicine studies due to the clinical applications that could derive from it. Dental pulp pluripotent stem cells are a considerable promise in tissue engineering and regenerative medicine as a source of tissue-specific cells, therefore the aim of this study is to demonstrate their ability to generate functional hepatocyte-like cells in vitro. Cells were cultured in a 2D environment and they were supplemented with several growth factors that recapitulate liver development. The expression of hepatic markers was detected by qRT-PCR and was confirmed by immunofluorescence analysis. The functional efficacy of the differentiated hepatic cells was evaluated by the secretion levels of albumin, cytochrome P450 activity, glycogen storage capacity and the production of hepatic enzymes was also evaluated. In this study we developed a 2D differentiation protocol of DPPSCs into hepatic lineage in vitro. The protocol was optimized to obtain definitive endoderm during the first days of differentiation and was continued with a modified five-step induction medium for 22 days based on liver development. Differentiated cells were able to express hepatic proteins during the entire differentiation process and were able to carry out several functions expected in mature hepatocytes. In this study we demonstrate that DPPSCs can be a powerful tool for liver regenerative medicine treatments to be developed in the future.