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J. Lipid Res.
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Journal of Lipid Research, Vol. 41, 882-893, June 2000
Copyright © 2000 by Lipid Research, Inc.

Original Article

An immortalized rat liver stellate cell line (HSC-T6): a new cell model for the study of retinoid metabolism in vitro

Silke Vogela, Roseann Piantedosia, Jorge Frankb, Avraham Lalazard, Don C. Rockeye, Scott L. Friedmand, and William S. Blanera,c
a Department of Medicine, College of Physicians and Surgeons of Columbia University, New York, NY, 10032
b Department of Dermatology, College of Physicians and Surgeons of Columbia University, New York, NY, 10032
c Institute of Human Nutrition, College of Physicians and Surgeons of Columbia University, New York, NY, 10032
d Division of Liver Diseases, Duke University Medical Center, Durham, NC 27710
e The Mount Sinai School of Medicine, New York, NY 10029: and Duke Liver Center, Duke University Medical Center, Durham, NC 27710

Correspondence to: William S. Blaner

Hepatocytes and hepatic stellate cells play important roles in retinoid storage and metabolism. Hepatocytes process postprandial retinyl esters and are responsible for secretion of retinol bound to retinol-binding protein (RBP) to maintain plasma retinol levels. Stellate cells are the body's major cellular storage sites for retinoid. We have characterized and utilized an immortalized rat stellate cell line, HSC-T6 cells, to facilitate study of the cellular aspects of hepatic retinoid processing. For comparison, we also carried out parallel studies in Hepa-1 hepatocytes. Like activated primary stellate cells, HSC-T6 express myogenic and neural crest cytoskeletal filaments. HSC-T6 cells take up and esterify retinol in a time- and concentration-dependent manner. Supplementation of HSC-T6 culture medium with free fatty acids (up to 300 µM) does not affect retinol uptake but does enhance retinol esterification up to 10-fold. RT-PCR analysis indicates that HSC-T6 cells express all 6 retinoid nuclear receptors (RAR{alpha}, -ß, -{gamma}, and RXR{alpha}, -ß, -{gamma}) and like primary stellate cells, HSC-T6 stellate cells express cellular retinol-binding protein, type I (CRBP) but fail to express either retinol-binding protein (RBP) or transthyretin (TTR). Addition of retinol (10-8;–10-5 M) or all-trans-retinoic acid (10-10;–10-6 M) rapidly up-regulates CRBP expression. Using RAR-specific agonists and antagonists and an RXR-specific agonist, we show that members of the RAR-receptor family modulate HSC-T6 CRBP expression.

Thus, HSC-T6 cells display the same retinoid-related phenotype as primary stellate cells in culture and will be a useful tool for study of hepatic retinoid storage and metabolism.—Vogel, S., R. Piantedosi, J. Frank, A. Lalazar, D. C. Rockey, S. L. Friedman, and W. S. Blaner. An immortalized rat liver stellate cell line (HSC-T6): a new cell model for the study of retinoid metabolism in vitro. J. Lipid Res. 2000. 41: 882;–893.

Supplementary key words: hepatic cells, Ito cells, fat-storing cells, vitamin A, oleic acid, cellular retinol-binding protein, type I (CRBP-I), retinol-binding protein (RBP), transthyretin (TTR), Hepa-1 hepatocytes, retinoid nuclear receptor


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