Dual phenotypic expression of hepatocytes and bile ductular markers in developing and preneoplastic rat liver

Abstract

This study supports the existence of a pluripotent liver stem cell population which has the potential to differentiate into hepatocytes and bile ductular cells. We compared the expression of hepatocyte-specific and bile ductular-specific markers in fetal and preneoplastic rat liver. L-pyruvate kinase (L-PK) and alpha glutathione S-transferase (GST) were used as adult hepatocyte-specific markers, while cytokeratin 19 (CK19) was used as a bile ductular-specific marker. pi GST and M2-pyruvate kinase (M2-PK), which are fetal hepatocyte-specific and expressed at high levels in the oval and duct-like cells, were also used. We characterized fetal liver derived from 13-21 days of gestation (E13-E21). pi GST was detected in the E18 hepatoblasts, which form the intrahepatic bile ducts, while CK19 was detected at E19. Some of these cells express alpha GST and L-PK from E19 to E21. Oval, duct-like and bile ductular cells in rats treated with a choline-deficient diet containing 0.07% ethionine (CDE diet) for up to 8 weeks were characterized by double immunocytochemistry. L-PK and alpha GST are absent from bile ductular cells in the normal adult liver and up to 3 weeks of CDE treatment. After 4-5 weeks on CDE treatment, the majority of bile ductular cells express L-PK, while at 6 weeks some co-express L-PK and alpha GST. There are two populations of oval cells, a major population expressing only the fetal hepatocyte markers, while a minor population expresses the fetal hepatocyte, adult hepatocyte and bile ductular markers. There are at least three different duct-like cell populations which co-express different markers and have characteristics of fetal hepatocytes at sequential stages of differentiation. One population co-expresses pi GST and M2-PK and is similar to fetal hepatocytes derived from E13-E14 fetuses. The second expresses the two fetal markers and L-PK, and this reflects characteristics of E15 hepatocytes. The third expresses pi GST, M2-PK, L-PK and alpha GST which is characteristic of E16-E19 hepatocytes. Upon withdrawal of the CDE diet, autoradiography using tritiated thymidine shows that oval and duct-like cells differentiate into hepatocytes. This study demonstrates that oval and duct-like cells express both hepatocytic and bile ductular markers, and have the capacity to differentiate into hepatocytes, characteristics similar to hepatoblasts in the developing rat liver.

Hedgehog-mediated mesenchymal–epithelial interactions modulate hepatic response to bile duct ligation

“…[73][74][75] Immunohistochemistry of BDL livers demonstrated that some bile ductular cells expressed Ptc and Gli-2, suggesting that relatively immature liver epithelial cells are Hh targets. Our in vitro studies support this concept because, under our culture conditions, the Ptc-( þ )ve cholangiocyte line coexpressed several oval cell markers, mpk and c-kit, [44][45][46][47] and the immature cholangiocyte marker, N-CAM, 3,43,76,77 as well as markers of more mature cholangiocytes (ie, aquaporin-1 and CK-19). If further research verifies that Hh signaling promotes the outgrowth of relatively immature liver epithelial cells, this might explain why patchy lobular necrosis was worse in PtcLacZ mice than WT mice after BDL.…”

“…45 The latter transcripts were also demonstrated in oval cells, which are known to coexpress neural, smooth muscle and progenitor markers. [44][45][46][47] These data suggest that Hh pathway activity occurs in immature cholangiocytes (ie, bile ductular cells).…”

Differential lymphotoxin-? and interferon gamma signaling during mouse liver regeneration induced by chronic and acute injury

“…More importantly, MPK staining is present in small hepatocytes, many of which also express the liver-specific isoenzyme of pyruvate kinase. 35 This finding suggests that A6 and MPK may distinguish oval cells that have differentiated along the bile duct and hepatocyte lineages, respectively. If this is correct, the finding that in IFN␥ KO mice only MPK ϩ oval cells are reduced suggests this cytokine affects the generation of hepatocytes, rather than bile duct cells, from oval cells.…”

Oval cell‐mediated liver regeneration: Role of cytokines and growth factors

“…It is our view that two distinct populations exist in rats fed a choline-deficient, ethionine-supplemented (CDE) diet, as detailed analysis of pyruvate kinase (PK) and GST isoenzyme expression reveals that most oval cells coexpress fetal and biliary specific markers, while a minor population coexpress fetal and adult hepatocyte markers. 9,10 Accordingly, oval cells are a multipotent cell population with the potential to differentiate into hepatocytes and biliary epithelia, and under certain conditions, pancreatic and intestinal epithelia (Fig. 1).…”