Transplantation of Embryonic Small Hepatocytes Induces Regeneration of Injured Liver in Adult Rat

Small hepatocytes as hepatic stem cells or progenitors may be transplanted to treat several end-stage liver diseases. To identify the characteristics of epithelial cells enriched from fetal liver, we used immunocytochemistry and electron micrography. All cells in the colonies were immunocytochemically positive for alpha fetoprotein and cytokeratins (CK) 7, CK8, and CK18, which are markers of hepatic progenitor. Under transmission electron microscopy, we observed the cultured cells to show naive characteristics of stem cells and to be significantly distinct from mature hepatocytes. To identity whether these small hepatocytes were able to proliferate and differentiate into mature hepatocytes, we cultured them in vitro, and, through the portal vein, and transplanted elements whose membrane were stained with red fluorescence using PKH26 linker dye, into the livers of CCl4-treated rats that had been subjected to two-thirds partial hepatectomy. Significant liver regeneration was observed 30 days later in rats that did or did not receive the cells. The livers of hepatocytes recipients showed sharper edges and smoother surfaces than the control group. Diffused cells labeled with red fluorescence were observed in the portal area, with branch-like red fluorescence in regions near portal areas of some lobules, suggesting that these elements were involved in the repair of liver lobules and differentiation into mature hepatocytes. Our results revealed that small hepatocytes not only have characteristics of hepatic stem cells, but also may be a source of cellular transplantation to treat liver diseases.