Reprogramming of human hepatocyte into bi-potent chemically derived hepatic progenitors

Abstract

Regenerative medicine is one of the most attainable and alternative breakthrough technologies to overcome the problems of organ shortages in transplantation. In case of chronic liver diseases, it is useful cell source for cell therapy that hepatocytes-like cells generated from patient fibroblasts by reprogramming into induced pluripotent stem cells, or by direct converting. However, these approaches are not sufficient to repopulate damaged liver. For this reason, we established a novel and stable induction method of patient specific hepatic progenitors from primary human hepatocytes using two independent small molecules, A83-01 and CHIR99021 and hepatocyte growth factor (HGF). Three days after treatment of two different small molecule agents and HGF to human primary hepatocytes, homogenous polygonal cells appear and grows. These human chemically derived hepatic progenitors (hCdHs) express hepatic and bile duct epithelial lineage genes and stained with hepatic progenitor markers. Next generation sequencing (NGS) study show rapid growing cells are closely related to human hepatoblasts. Also, hCdHs differentiate into hepatocytes and biliary epithelial cells suggesting property of bipotent stem cells. After 10 passages, hCdHs doesn’t lose growth pattern and hepatic progenitor phenotype. hCdHs also repopulate in the immunocompromised mouse model after transplantation through intrasplenic route. In conclusion, hCdHs, which is bipotent progenitor cells, is easily induced by small molecules, and effectively recover damaged liver without organoid formation. Thus, hCdHs induction technique could open a new era of field for regenerative medicine of the liver.