Development of dexamethasone‐inducible tyrosine aminotransferase activity in WB‐F344 rat liver epithelial stemlike cells cultured in the presence of sodium butyrate

Abstract

Sodium butyrate acts as a differentiation-promoting agent for a wide variety of cell types, including some tumor cell lines. In this study, we examined the effects of sodium butyrate (SB) on the functional differentiation of cultured WB-F344 rat liver epithelial stemlike cells. Treatment of WB-F344 cells with 3.75 mM SB resulted in an inhibition of cellular proliferation, alterations to normal cellular morphology (increased cell size and decreased nuclear/cytoplasmic ratio), and significant increases in cellular protein synthesis. The SB-mediated changes in cell morphology, proliferative status, and protein catabolism were accompanied by development of dexamethasone-inducible tyrosine aminotransferase (TAT) enzyme activity. Culture of WB-F344 cells in growth medium containing SB and dexamethasone (DEX; 1 x 10(-6) M) resulted in greater than sevenfold increase in the basal TAT activity compared with control cultures. An additional sixfold increase in TAT activity was observed when cells cultured in medium containing SB and DEX were exposed to 1 x 10(-7) M DEX during the last 24 hours of culture. The DEX-inducible TAT activity developed by SB-treated WB-F344 cells responded to the modulating effects of insulin and L-tyrosine in a manner that closely resembled that reported for cultured hepatocytes and hepatoma cell lines. These studies show that treatment of WB-F344 rat liver epithelial stemlike cells with the differentiation-promoting agent SB in vitro leads to expression of the differentiation-specific hepatocyte enzyme TAT.

Epithelial cells with hepatobiliary phenotype: Is it another stem cell candidate for healthy adult human liver?

“…Such phenomenon has been documented for porcine LECs [18,27] . Nevertheless, other studies demonstrated the ability of LECs to differentiate into hepatocytes and biliary cells after incubation with growth factors defined media [28,29] . In the rat model, normal LECs have also been shown to engraft and to differentiate into hepatocytes or biliary cells [30,31] .…”

Expression of matrix metalloproteinase-2 and -9 and of tissue inhibitor of matrix metalloproteinase-1 in liver regeneration from oval cells in rat

“…We also studied OSM-R expression by WB-F344 cells treated or not with the association of sodium butyrate and dexamethasone. This treatment is known to induce hepatocytic differentiation of WB-F344 cells (Coleman et al, 1994), as revealed by an increased production of albumin (Fig. 6c).…”

“…In vitro, these cells exhibit a poorly differentiated epithelial phenotype. After exposure to differentiation stimuli, they express phenotypic characteristics of hepatocytes (Coleman et al, 1994) or biliary epithelial cells (Couchie et al, 2002) and acquire morphological and functional properties of hepatocytes when injected into the liver of syngenic animals (Coleman et al, 1997). Zymographic analysis of conditioned medium from untreated WB-F344 cells revealed that these cells synthesized MMP-2 and MMP-9 in their proform (66 and 95 kDa) (Fig.…”

“…Cells were routinely cultured in Ham's F12 medium (Invitrogen, Cergy Pontoise, France) supplemented with 5% fetal calf serum (FCS) (complete medium). WB-F344 differentiation towards the hepatocytic lineage was induced as previously described (Coleman et al, 1994). In brief, 5 × 10 5 cells were seeded in 10-cm dishes and allowed to adhere for 24 h. Cells were incubated for 6 days in complete medium in the absence (untreated) or in the presence (treated) of 3.75 mmol/l sodium butyrate and 1 μmol/l dexamethasone.…”

Fluorescently Labeled Nanoparticles Enable the Detection of Stem Cell-Derived Hepatocytes

“…Tyrosine aminotransferase (TAT) is a wellknown enzymatic marker for hepatocyte-specific differentiation. 27 Frankly, we failed to amplify the human TAT gene even when we used nested RT-PCR from cDNA made from whole bovine liver. As in Figure 1, SPIO-containing hMSCs were seldom detected in liver tissue.…”