Isolation of hepatoblasts based on the expression of Dlk/Pref-1

摘要: Hepatoblasts are common progenitors for hepatocytes and biliary epithelial cells, although their nature remains largely unknown. In order to isolate characterize hepatoblasts, we searched cell surface antigens expressed in mouse fetal hepatic cells by the signal sequence trap method found that Dlk, also known as Pref-1, was strongly liver. Immunohistochemical well northern analysis indicated Dlk highly E10.5 liver bud. The strong expression continued until E16.5 stage significantly downregulated thereafter. Using a monoclonal antibody against isolated Dlk+ either fluorescence-activated sorter or an automatic magnetic sorter. from livers albumin formed colonies when cultured at low density with HGF EGF 5 days. Over 60% of derived E14.5 contained both albumin+ cytokeratin 19+ indicating majority colony-forming able differentiate into hepatocyte lineages. addition, numerous microvilli were observed electronmicroscopic most those differentiation under this condition. Furthermore, 7% not only bipotential but proliferative, forming large colony containing more than 100 during days culture. By transplantation spleen, donor-derived recipient liver, differentiated vivo. These results indicate hepatoblasts is useful marker enrich proliferative