Flow cytometric isolation and clonal identification of self-renewing bipotent hepatic progenitor cells in adult mouse liver

Abstract

The adult liver progenitor cells appear in response to several types of pathological liver injury, especially when hepatocyte replication is blocked. These cells are histologically identified as cells that express cholangiocyte markers and proliferate in the portal area of the hepatic lobule. Although these cells play an important role in liver regeneration, the precise characterization that determines these cells as self-renewing bipotent primitive hepatic cells remains to be shown. Here we attempted to isolate cells that express a cholangiocyte marker from the adult mouse liver and perform single cell-based analysis to examine precisely bilineage differentiation potential and self-renewing capability of these cells. Based on the results of microarray analysis and immunohistochemistry, we used an antibody against CD133 and isolate CD133 ؉ cells via flow cytometry. We then cultured and propagated isolated cells in a single cell culture condition and examined their potential for proliferation and differentiation in vitro and in vivo. Isolated cells that could form large colonies (LCs) in culture gave rise to both hepatocytes and cholangiocytes as descendants, while maintaining undifferentiated cells by self-renewing cell divisions. The clonogenic progeny of an LCforming cell is capable of reconstituting hepatic tissues in vivo by differentiating into fully functional hepatocytes. Moreover, the deletion of p53 in isolated LC-forming cells resulted in the formation of tumors with some characteristics of hepatocellular carcinoma and cholangiocarcinoma upon subcutaneous injection into immunodeficient mutant mice. These data provide evidence for the stem cell-like capacity of isolated and clonally cultured CD133 ؉ LC-forming cells. Conclusion: Our method for prospectively isolating hepatic progenitor cells from the adult mouse liver will facilitate study of their roles in liver regeneration and carcinogenesis. (HEPATOLOGY 2008;48

FGF7 is a functional niche signal required for stimulation of adult liver progenitor cells that support liver regeneration

“…It has been well documented that stem/progenitor cell activity of a certain population of liver cells can be defined by their capacity to generate large colonies that are capable of expressing both hepatocyte and BEC lineage markers in culture (Suzuki et al 2008;Okabe et al 2009;Dorrell et al 2011;Shin et al 2011). When EpCAM + cells isolated from the FGF7 Tg mice and the control mice were subjected to in vitro colony formation assays (Okabe et al 2009), the EpCAM + cells from the Tg mice formed colonies, including those composed of >100 cells (Fig.…”

Simultaneous characterization of progenitor cell compartments in adult human liver

“…2, panel C). Moreover, freshly isolated L-EpPCs expressed epithelial cell adhesion molecule (EpCAM) and CD133-1, which have recently been identified as markers of hepatic progenitor cells (20,21,23), but this expression progressively decreased in culture, whereas the expression of neural cell adhesion molecule (NCAM) and CD166 antigens increased during the five culture passages (Fig. 2, panel C).…”

Cancer Stem Cells and Aldehyde Dehydrogenase 1 in Liver Cancers

“…During serious liver injury in rodents, the number of characteristic nonparenchymal oval cells increases in the periportal regions. These cells express both cholangiocellular (Ck7 and Ck19) and hepatocellular (Afp and Alb) marker genes and diferentiate into both hepatocytic and cholangiocytic cells, suggesting that oval cells are candidate hepatic progenitors [20][21][22][23].…”