Continuous epithelial cell lines from ADPKD liver cysts exhibit characteristics of intrahepatic biliary epithelium

Abstract

We have produced continuous cell lines using retroviral transduction of SV40 large T antigen into epithelial cells removed from the lumen of liver cysts from four female patients with autosomal dominant polycystic kidney disease (ADPKD). Liver cyst-derived epithelial (LCDE) cell lines are grown in a hormonally supplemented medium in the presence of lethally irradiated NIH/3T3 fibroblast coculture. LCDE cells maintain their epithelial appearance and are positive for the biliary-specific markers cytokeratin 7 and 19 and gamma-glutamyl transpeptidase while being negative for hepatocyte markers. SV40 large T antigen is localized to the cell nucleus. LCDE cells have been grown continuously for periods exceeding 12 mo and 25 passages (170 population doublings). LCDE cells exhibit intracellular pH regulatory pathways that, with one exception, are similar to those found in normal intrahepatic biliary epithelium. These LCDE cell lines exhibit impaired alkalinization in response to Cl- substitution. This finding is suggestive of decreased function or abundance of a Cl-/HCO3- anion exchanger and could account for the failure of ADPKD hepatic cysts to secrete HCO3- in response to secretin.

Morphological and Functional Features of Hepatic Cyst Epithelium in Autosomal Dominant Polycystic Kidney Disease

“…Although less than 1% of cholangiocytes lining intrahepatic bile ducts of 20 to 100 m displayed a thin positivity for pAKT ( Figure 5A), the epithelium lining hepatic cysts showed an intense and diffuse cytoplasmic and nuclear pAKT positivity (Figure 5B) suggesting activation of the IGF1 system that together with estrogens probably sustains the enhanced proliferative activities characterizing cyst epithelium. [5][6][7] In fact, the cyst epithelial cells showed an intense staining for PCNA ( Figure 5D), a marker of cell proliferation, involving almost all cells, whereas in the normal intrahepatic biliary epithelium PCNA was virtually negative ( Figure 5C), consistent with the quiescent status of normal cholangiocytes. 10 …”

“…[1][2][3][4] Hepatic cysts are lined by epithelial cells featuring phenotypical and functional characteristics of biliary epithelium with enhanced secretory and proliferative activities. [5][6][7] Fluid secretion, extracellular matrix remodeling, proliferation of the lining epithelial cells, and neovascularization each are considered important steps in promoting cyst growth. 8 Very recently, a number of growth factors and cytokines that are increased in serum or cystic fluid have been shown to drive cyst growth and expansion.…”

Repeated Minocycline Hydrochloride Injections for Symptomatic Polycystic Liver Disease

“…Hemorrhagic or infected cysts were excluded from this study. Exclusion criteria were contraindications to percutaneous treatment (a platelet count less than 50,000/mm 3 ) and the concomitant presence of hepatobiliary malignancy, hemorrhagic cyst, or infected cyst. Percutaneous fluid sampling was performed as required for cytological and chemical analysis.…”

“…Typically, disconnected biliary structures are present in veryearly-stage disease [1] . Because the hepatic cyst epithelium shows the immunohistochemical, structural, and functional features of cholangiocytes, a hepatic cyst can be described as a fluid-filled cavity lined with cholangiocytes [2][3][4][5] . Most patients with PLD remain asymptomatic, but 2% to 5% of patients will have symptomatic hepatomegaly caused by a continuous increase in the volumes cysts.…”

Surgical management of polycystic liver disease

“…The growth of cysts in the liver is thought to arise from cell proliferation, solute and fluid secretion into the cysts, and expansion of abnor mal cell matrices. Perrone and colleagues demonstrated, via culture derived epithelial cell lines, that these cysts are of biliary origin [7] . Morphological studies demonstrate that the peripheral cysts arise from biliary microhamartomas, but the centrally located cysts arise from dilatation of the peribiliary glands in the liver [8] .…”