Development of a Non-Transformed Human Liver Cell Line with Differentiated-Hepatocyte and Urea-Synthetic Functions: Applicable for Bioartificial Liver

Abstract

There is a need to develop human hepatocyte cell lines which retain both replicating capacity and highly differentiated functions to facilitate the development of an efficient bioartificial liver. The present study was undertaken to differentiate, using sodium butyrate, the actively replicating immortalized human liver cell line. The effects of butyrate on cell growth and cell cycle were analyzed, and the albumin synthesis, cytochrome P450 and ammonia-detoxifying activity of the butyrate-treated cells were measured. Butyrate treatment resulted in G2/M arrest of the cell cycle and polygonal changes in the cell morphology. Neither the control nor the butyrate-treated cells showed transformed characteristics. Butyrate treatment increased the amount of albumin secretion, cytochrome P450 activity, and the urea production rate of the cells. The present study provides non-transformed human hepatocytes, which can replicate unlimitedly and then restore differentiated hepatocyte-specific functions by butyrate, and therefore, have applications for the development of an efficient bioartificial liver.

Multipotent adult progenitor cells from bone marrow differentiate into functional hepatocyte-like cells

“…However, the degree of differentiation measured by absence of immature markers such as αFP and CK19 was higher when cells were also treated with HGF. This is consistent with the observation that FGF-4 is important in initial endoderm patterning and may play a role in endoderm specification (23) and that HGF induces differentiation of hepatocytes that are not actively proliferating (28)(29)(30)(31). Differentiation to cells with a hepatocyte phenotype was seen only when cells were seeded at high confluence, which may cause a postmitotic state required for differentiation in the presence of HGF.…”

Ursodeoxycholic acid-induced inhibition of DLC1 protein degradation leads to suppression of hepatocellular carcinoma cell growth

“…cells were grown in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum, 100,000 U/liter penicillin and 100 mg/l streptomycin at 37˚C in a humidified atmosphere of 5% CO 2 and 95% air. The immortalized human hepatocytes (15) were maintained over 150 passages in a chemically defined Ham's F-12 medium (sigma chemical co., st. louis, Mo) containing 50 ng/ml epidermal growth factor (collaborative Biomedical, Waltham, MA), 10 µg/ml insulin, 5 μg/ml transferring, 3.5 µg/M hydrocortisone, 2 mM glutamine, 0.1 µM cuso 4 • 5H 2 o, 0.03 µM H 2 seo 3 , 0.5 nM Znso 4 • 7H 2 o, 1 ng/ml glucagon, 6.5 ng/ml growth hormone and 0.5 µg/ ml linoleic acid (sigma chemical co.). we used UDcA, which is a secondary hydrophilic bile acid, and chenodeoxycholate (cDc), a hydrophobic bile acid, for this study (sigma chemicals co.).…”

Recent advances in re-engineered liver: de-cellularization and re-cellularization techniques

“…Hepatoblasts, human fetal hepatocytes and ESCs may hold promise for the future because of their potential for unlimited cellular expansion. Fetal hepatocytes have been immortalized by SV40 T antigen [77] or human telomerase reverse transcriptase [78]. Moreover, human fetal liver cell transplantation has been applied in the treatment of chronic liver disease [79], and the infusion of fetal liver cells is a safe and welltolerated procedure in patients [79].…”