Re-maturation of canine chemically-induced hepatic progenitor cells toward biliary epithelial cells

Abstract

This study aimed to evaluate the differentiation potential of canine hepatic progenitor cells (cHPCs) derived from cryopreserved hepatocytes (cHeps). The cHeps were reprogrammed using three small molecules, Y-27632, A-83-01, and CHIR99021, to establish cHPCs. The cells were cultured on Matrigel in a chemically defined feeder-free maintenance medium for human pluripotent stem cells containing YAC, followed by 2% growth factor-reduced Matrigel. During culture, cHPCs transitioned from two-dimensional sheets to three-dimensional ductal structures. qRT-PCR and immunohistochemistry revealed increased CK19 expression on the ductal surfaces, indicating biliary epithelial-like differentiation. Overall, cHPCs possess a bipotential rematuration capacity. This study demonstrated that cHPCs induced by the YAC cocktail possess bipotent differentiation capabilities, confirming that their characteristics are likely to chemically induce liver progenitor cells.