Immortal hepatic stellate cell lines: Useful tools to study hepatic stellate cell biology and function?

Journal of Cellular and Molecular Medicine

Volumn 11, Issue 4, 2007, Pages 704-722

  Herrmann, Jens ^a   Gressner, Axel M ^a   Weiskirchen, Ralf ^a  

^a UNIVERSITY HOSPITAL RWTH AACHEN   (Germany)

Author keywords

Cell culture; Hepatic fibrogenesis; Hepatic stellate cells; Immortalization; Liver

Indexed keywords

ANIMAL; CELL LINE; CULTURE TECHNIQUE; CYTOLOGY; HUMAN; LIVER CELL; METABOLISM; METHODOLOGY; REVIEW;

ANIMALS; CELL CULTURE TECHNIQUES; CELL LINE, TRANSFORMED; HEPATOCYTES; HUMANS;

ANIMALIA; RODENTIA;

EID: 34548180280     PISSN: 15821838     EISSN: None     Source Type: Journal    
DOI: 10.1111/j.1582-4934.2007.00060.x     Document Type: Review

References (84)

1. De Leeuw AM, McCarthy SP, Geerts A, Knook DL. Purified rat liver fat-stroing cells in culture divide and contain collagen. Hepatology. 1984 4 : 392 403.
2. Weiskirchen R, Gressner AM. Isolation and culture of hepatic stellate cells. Methods Mol Med. 2005 117 : 99 113.
3. Geerts A, Niki T, Hellemans K, De Craemer D, Van Den Berg K, Lazou JM, Stange G, Van De Winkel M, De Bleser P. Purification of rat hepatic stellate cells by side scatter-activated cell sorting. Hepatology. 1998 27 : 590 8.
4. Friedman SL, Rockey DC, McGuire RF, Maher JJ, Boyles JK, Yamasaki G. Isolated hepatic lipocytes and Kupffer cells from normal human liver: morphological and functional characteristics in primary culture. Hepatology. 1992 15 : 234 43.
5. Blazejewski S, Preaux AM, Mallat A, Brocheriou I, Mavier P, Dhumeaux D, Hartmann D, Schuppan D, Rosenbaum J. Human myofibroblastlike cells obtained by outgrowth are representative of the fibrogenic cells in the liver. Hepatology. 1995 22 : 788 97.
6. Kmiec Z. Cooperation of liver cells in health and disease. Adv Anat Embryol Cell Biol. 2001 161 : 1 151.
7. Geerts A. History, heterogeneity, developmental biology, and functions of quiescent hepatic stellate cells. Semin Liver Dis. 2001 21 : 311 35.
8. Gressner AM, Weiskirchen R. Modern pathogenetic concepts of liver fibrosis suggest stellate cells and TGF-_ as major players and therapeutic targets. J Cell Mol Med. 2006 10 : 76 99.
9. Herbst H, Frey A, Heinrichs O, Milani S, Bechstein WO, Neuhaus P, Schuppan D. Heterogeneity of liver cells expressing procollagen types I and IV in vivo. Histochem Cell Biol. 1997 107 : 399 409.
10. Zou Z, Ekataksin W, Wake K. Zonal and regional differences identified from precision mapping of vitamin A-storing lipid droplets of the hepatic stellate cells in pig liver: a novel concept of addressing the intralobular area of heterogeneity. Hepatology. 1998 27 : 1098 108.
11. Knittel T, Kobold D, Saile B, Grundmann A, Neubauer K, Piscaglia F, Ramadori G. Rat liver myofibroblasts and hepatic stellate cells: different cell populations of the fibroblast lineage with fibrogenic potential. Gastroenterology. 1999 117 : 1205 21.
12. Magness ST, Bataller R, Yang L, Brenner DA. A dual reporter gene transgenic mouse demonstrates heterogeneity in hepatic fibrogenic cell populations. Hepatology. 2004 40 : 1151 9.
13. Kordes C, Sawitza I, Müller-Marbach A, Ale-Agha N, Keitel V, Klonowski-Stumpe H, Häussinger D. CD133+ hepatic stellate cells are progenitor cells. Biochem Biophys Res Commun. 2007 352 : 410 7.
14. Kristensen DB, Kawada N, Imamura K, Miyamoto Y, Tateno C, Seki S, Kuroki T, Yoshizato K. Proteome analysis of rat hepatic stellate cells. Hepatology. 2000 32 : 268 77.
15. Borojevic R, Monteiro AN, Vinhas SA, Domont GB, Mourao PA, Emonard H, Grimaldi G Jr., Grimaud JA. Establishment of a continuous cell line from fibrotic schistosomal granulomas in mice livers. In vitro Cell Dev Biol. 1985 21 : 382 90.
16. Guma FCR, Mello TG, Mermelstein CS, Fortuna VA, Wofchuk ST, Gottfried C, Guaragna RM, Costa ML, Borojevic R. Intermediate filaments modulation in an in vitro model of the hepatic stellate cell activation or conversion into the lipocyte phenotype. Biochem Cell Biol. 2001 79 : 409 17.
17. Mermelstein CS, Guma FC, Mello TG, Fortuna VA, Guaragna RM, Costa ML, Borojevic R. Induction of the lipocyte phenotype in murine hepatic stellate cells: reorganisation of the actin cytoskeleton. Cell Tissue Res. 2001 306 : 75 83.
18. Horie S, Kitamura Y, Kawasaki H, Terada T. Inhibitory effects of antisense oligonucleotides on the expression of procollagen type III gene in mouse hepatic stellate cells transformed by simian virus 40. Pathol Int. 2000 50 : 937 44.
19. Kitamura Y, Tanigawa T, Katsumoto T, Tomita K, Wang HR, Hirai K, Ichihara K, Terada T. Cell growth and differentiation of a novel mouse Ito (fat-storing) cell line transformed by a temperature-sensitive mutant of simian virus 40. Hepatology. 1997 26 : 323 9.
20. Proell V, Mikula M, Fuchs E, Mikulits W. The plasticity of p19 ARF null hepatic stellate cells and the dynamics of activation. Biochim Biophys Acta. 2005 1744 : 76 87.
21. Matsuura T, Kawada M, Sujino H, Hasumura S, Nagamori S, Shimizu H. Vitamin A metabolism of immortalized hepatic stellate cells in the bioreactor. In : Wisse E, Knook DL, De Zanger R, Arthur MJP, editors. Cells of the Hepatic Sinusoid 7. Leiden : Kupffer Cell Foundation 1999. pp. 88 89.
22. Vogel S, Piantedosi R, Frank J, Lalazar A, Rockey DC, Friedman SL, Blaner WS. 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 93.
23. Greenwel P, Schwartz M, Rosas M, Peyrol S, Grimaud JA, Rojkind M. Characterization of fatstoring cell lines derived from normal and CCl4-cirrhotic livers. Differences in the production of interleukin- 6. Lab Invest. 19991 65 : 644 53.
24. Greenwel P, Rubin J, Schwartz M, Hertzberg EL, Rojkind M. Liver fat-storing cell clones obtained from a CCl4-cirrhotic rat are heterogeneous with regard to proliferation, expression of extracellular matrix components, interleukin-6, and connexin 43. Lab Invest. 1993 69 : 210 6.
25. Sauvant P, Sapin V, Abergel A, Schmidt CK, Blanchon L, Alexandre-Gouabau MC, Rosenbaum J, Bommelaer G, Rock E, Dastugue B, Nau H, Azais-Braesco V. PAV-1, a new rat hepatic stellate cell line converts retinol into retinoic acid, a process altered by ethanol. Int J Biochem Cell Biol. 2002 34 : 1017 29.
26. Sauvant P, Abergel A, Partier A, Alexandre-Gouabau MC, Rock E, Sion B, Motta C, Sapin V, Azais-Bresco V. Treatment of the rat hepatic stellate cell line, PAV-1, by retinol and palmitic acid leads to a convenient model to study retinoids metabolism. Biol Cell. 2002 94 : 401 8.
27. Pan Q, Li DG, Lu HM, Wang YQ, Zhang WZ, Xu QF. A new immortalized rat cell line, hepatic stellate cell-PQ, exhibiting characteristics of hepatic stellate cell. Hepatobiliary Pancreat Dis Int. 2005 4 : 281 4.
28. Xiong S, Yavrom S, Hazra S, Wu D, She H. Spontaneously immortalized cell line from rat liver fibrosis with phenotypes of both hepatic stellate cells and myofibroblasts. Hepatology 2001 34 : 520A.
29. Sung SK, She H, Xiong S, Tsukamoto H. Tumor necrosis factor-_ inhibits peroxisome proliferator-activated receptor _ activity at a posttranslational level in hepatic stellate cells. Am J Physiol Gastrointest Liver Physiol. 2003 286 : G722 9.
30. Isono M, Soda M, Inoue A, Akiyoshi H, Sato K. Reverse transformation of hepatic myofibroblast-like cells by TGF_1/LAP. Biochem Biophys Res Commun. 2003 311 : 959 65.
31. Kim JY, Kim KM, Nan JX, Zhao YZ, Park PH, Lee SJ, Sohn DH. Induction of apoptosis in transhinone I via cytochrome c release in activated hepatic stellate cells. Pharmacol & Toxicol. 2003 92 : 195 200.
32. Tahashi Y, Matsuzaki K, Date M, Yoshida K, Furukawa F, Sugano Y, Matsushita M, Himeno Y, Inagaki Y, Inoue K. Differential regulation of TGF-_ signal in hepatic stellate cells between acute and chronic rat liver injury. Hepatology. 2002 35 : 49 61.
33. Rao HY, Wei L, Fei R, Wang JH, Jiang D, Zhang Q, Cong X. Inhibitory effect of interferon-_ on the activation of LX-2 and rHSC-99 hepatic stellate cells in culture. Zhonghua Gan Zang Bing Za Zhi. 2006 14 : 550 2.
34. Murakami K, Abe T, Miyazawa M, Yamaguchi M, Masuda T, Matsuura T, Nagamori S, Takeuchi K, Abe K, Kyogoku M. Establishment of a new human cell line, LI90, exhibiting characteristics of hepatic Ito (fat-storing) cells. Lab Invest. 1995 72 : 731 9.
35. Shibata N, Watanabe T, Okitsu T, Sakaguchi M, Takesue M, Kunieda T, Omoto K, Yamamoto S, Tanaka N, Kobayashi N. Establishment of an immortalized human hepatic stellate cell line to develop antifibrotic therapies. Cell Transplant. 2003 12 : 499 507.
36. Watanabe T, Shibata N, Westerman KA, Okitsu T, Allain JE, Sakaguchi M, totsugawa T, Maruyama M, Matsumura T, Noguchi H, Yamamoto S, Hikida M, Ohmori A, Reth M, Weber A, Tanaka N, Leboulch P, Kobayashi N. Establishment of immortalized human hepatic stellate scavenger cells to develop bioartificial livers. Transplantation. 2003 75 : 1873 80.
37. Weill FX, Blazejewski S, Blanc JF, Huet S, Gauthier JM, Neaud V, Olaso E, Dubuisson L, Azais-Braesco V, Vidal-Vanaclocha F, Balabaud C, Bioulac-Sage P, Rosenbaum J. Characterization of a new human liver myofibroblast cell line: transcriptional regulation of plasminogen activator inhibitor type I by transforming growth factor beta 1. Lab Invest. 1997 77 : 63 70.
38. Schnabl B, Choi YH, Olsen JC, Hagedorn CH, Brenner DA. Immortal activated human hepatic stellate cells generated by ectopic telomerase expression. Lab Invest. 2002 82 : 323 33.
39. Xu L, Hui AY, Albanis E, Arthur MJ, O'Byrne SM, Blaner WS, Mukherjee P, Friedman SL, Eng FJ. Human hepatic stellate cell lines, LX-1 and LX-2: new tools for analysis of hepatic fibrosis. Gut. 2005 54 : 142 51.
40. González-Cuevas J, Bueno-Topete M, Armendariz-Borunda. Urokinase plasminogen activator stimulates function of active forms of stromelysin and gelatinases (MMP-2 and MMP-9) in cirrhotic tissue. J Gastroenterol Hepatol 2006 21 : 1544 54.
41. Schnabl B, Purbeck CA, Choi YH, Hagedorn CH, Brenner D, Schnabl B, Purbeck CA, Choi YH, Shibata N, Watanabe T, Okitsu T, Sakaguchi M, Takesue M, Kunieda T, Omoto K, Yamamoto S, Tanaka N, Kobayashi N. Establishment of an immortalized human hepatic stellate cell line to develop antifibrotic therapies. Cell Transplant. 2003 12 : 499 507.
42. Schnabl B, Purbeck CA, Choi YH, Hagedorn CH, Brenner D. Replicative senescence of activated human hepatic stellate cells is accompanied by a pronounced inflammatory but less fibrogenic phenotype. Hepatology. 2003 37 : 653 64.
43. Friedman SL. Molecular regulation of hepatic fibrosis, an integrated cellular response to tissue injury. J Biol Chem. 2000 275 : 2247 50.
44. Gressner AM, Weiskirchen R, Breitkopf K, Dooley S. Roles of TGF-_ in hepatic fibrosis. Front Biosci. 2002 7 : d793 807.
45. Senoo H, Imai K, Sato M, Kojima N, Miura M, Hata RI. Three-dimensional structure of extracellular matrix reversibly regulates morphology, proliferation, and collagen metabolism of perisinusoidal stellate cells (Vitamin A-storing cells). Cell Biol Int. 1996 20 : 501 12.
46. Sohara N, Znoyko I, Levy MT, Trojanowska M, Reuben A. Reversal of activation of human myofibroblast- like cells by culture on a basement membrane- like substrate. J Hepatol. 2002 37 : 214 21.
47. Weiskirchen R, Kneifel J, Weiskirchen S, van de Leur E, Kunz D, Gressner AM. Comparative evaluation of gene delivery devices in primary cultures of rat hepatic stellate cells and rat myofibroblasts. BMC Cell Biol. 2000 1 : 4.
48. Ueki N, Ohkawa T, Yamamura H, Takahashi K, Tsutsui T, Kawai Y, Yokoyama Y, Amuro Y, Hada T, Higashino K. Induction of calponin-h1 by transforming growth factor-_1 in cultured human Ito cells, LI90. Biochim Biophys Acta. 1998 1403 : 28 36.
49. Tang M, Potter JJ, Mezey E. Leptin enhances the effect of transforming growth factor _ in increasing type I collagen formation. Biochem Biophys Res Commun. 2002 297 : 906 11.
50. Dooley S, Delvoux B, Lahme B, Mangasser-Stephan K, Gressner AM. Modulation of transforming growth factor beta response and signaling during transdifferentiation of rat hepatic stellate cells to myofibroblasts. Hepatology. 2000 31 : 1094 106.
51. Dooley S, Delvoux B, Streckert M, Bonzel L, Stopa M, ten Dijke P, Gressner AM. Transforming growth factor beta signal transduction in hepatic stellate cells via Smad2/3 phosphorylation, a pathway that is abrogated during in vitro progression to myofibroblasts. TGF_ signal transduction during transdifferentiation of hepatic stellate cells. FEBS Lett. 2001 502 : 4 10.
52. Pinzani M, Gesualdo L, Sabbah GM, Abboud HE. Effects of platelet-derived growth factor and other polypeptide mitogens on DNA synthesis and growth of cultured rat liver fat-storing cells. J Clin Invest. 1989 84 : 1786 93.
53. Borkham-Kamphorst E, van Roeyen CR, Ostendorf T, Floege J, Gressner AM, Weiskirchen R. Pro-fibrogenic potential of PDGF-D in liver fibrosis. J Hepatol. 2007 46 : 1064 74.
54. Bachem MG, Riess U, Gressner AM. Liver fat-storing cell proliferation is stimulated by epidermal growth factor/transforming growth factor β and inhibited by transforming growth factor β. Biochem Biophys Res Commun. 1989 162 : 708 14.
55. Westhoff JH, Sawitza I, Keski-Oja J, Gressner AM, Breitkopf K. PDGF-BB induces expression of LTBP-1 but not TGF-β 1 in a rat cirrhotic fat storing cell line. Growth Factors. 2003 21 : 121 30.
56. Inagaki Y, Truter S, Greenwel P, Rojkind M, Unoura M, Kobayashi K, Ramirez F. Regulation of the alpha 2(I) collagen gene transcription in fat-storing cells derived from a cirrhotic liver. Hepatology. 1995 22 : 573 9.
57. Garcia-Trevijano ER, Iraburu MJ, Fontana L, Dominguez-Rosales JA, Auster A, Covarrubias-Pinedo A, Rojkind M. Transforming growth factor beta1 induces the expression of alpha1(I) procollagen mRNA by a hydrogen peroxide-C/ EBPbetadependent mechanism in rat hepatic stellate cells. Hepatology. 1999 29 : 960 70.
58. Herrmann J, Borkham-Kamphorst E, Haas U, Van de Leur E, Fraga MF, Esteller M, Gressner AM, Weiskirchen R. The expression of CSRP2encoding the LIM domain protein CRP2 is mediated by TGF-_ in smooth muscle and hepatic stellate cells. Biochem Biophys Res Commun. 2006 345 : 1526 35.
59. Zhang YP, Yao XX, Zhao X. Interleukin-1 _ up-regulates tissue inhibitor of matrix metalloproteinase-1 mRNA and phosphorylation of c-jun N-terminal kinase and p38 in hepatic stellate cells. World J Gastroenterol. 2006 12 : 1392 6.
60. Inagaki Y, Nemoto T, Kushida M, Sheng Y, Higashi K, Ikeda K, Kawada N, Shirasaki F, Takehara K, Sugiyama K, Fujii M, Yamauchi H, Nakao A, de Crombrugghe B, Watanabe T, Okazaki I. Interferon alfa down-regulates collagen gene transcription and suppresses experimental hepatic fibrosis in mice. Hepatology. 2003 38 : 890 9.
61. Iraburu MJ, Dominguez-Rosales JA, Fontana L, Auster A, Garcia-Trevijano ER, Covarrubias-Pinedo A, Rivas-Estilla AM, Greenwel P, Rojkind M. Tumor necrosis factor alpha down-regulates expression of the alpha1(I) collagen gene in rat hepatic stellate cells through a p20 C/EBPbeta- and C/EBPdelta-dependent mechanism. Hepatology. 2000 31 : 1086 93.
62. Zhang M, Zhang JP, Ji HT, Wang JS, Qian DH. Effect of six flavonoids on proliferation of hepatic stellate cells in vitro. Acta Pharmacol Sin. 2000 21 : 253 6.
63. Song YH, Zhou XM, Xue XN, Liu NZ, Tian DA, Kong XJ, Wu XL, Lin JS, Jin YX. Effect of ribozyme against transforming growth factor _1 on biological character of activated HSCs. IUBMB Life. 2005 57 : 31 9.
64. Liu X, Wang W, Hu H, Tang N, Zhang C, Liang W, Wang M. Smad3 specific inhibitor, naringenin, decreases the expression of extracellular matrix induced by TGF-beta1 in cultured rat hepatic stellate cells. Pharm Res. 2006 23 : 82 9.
65. Gui M, Zhang YF, Xiao ZY, Sun P, Dai JF, Wang SF, Rui YC, Zhang JP. Inhibitory effect of emodin on tissue inhibitor of metalloproteinases-1 (TIMP-1) expression in rat hepatic stellate cells. Dig Dis Sci. 2007 52 : 200 7.
66. Kariv R, Enden A, Zvibel I, Rosner G, Brill S, Shafritz DA, Halpern Z, Oren R. Triiodothyronine and interleukin-6 (IL-6) induce expression of HGF in an immortalized rat hepatic stellate cell line. Liver Int. 2003 23 : 187 93.
67. Ma H, Ma XM, Yin CH, Jia JD, Wang BE. Effects of gamma-interferon on gene expression of collagen I, III and on the tissue inhibitor of metalloprotenase 1 in HSC-T6 cells. Zhonghua Gan Zang Bing Za Zhi. 2005 13 : 528 30.
68. Maubach G, Lim MC, Kumar S, Zhuo L. Expression and upregulation of cathepsin S and other early molecules required for antigen presentation in activated hepatic stellate cells upon IFN-gamma treatment. Biochim Biophys Acta. 2007 1773 : 219 31.
69. Li X, Meng Y, Cai SX, Yang XS, Zhang YJ, Wu PS. Angiotensin II and aldosterone stimulate alpha1-(I) procollagen mRNA expression in hepatic stellate cells via activation of ERK1/2 and AP-1. Zhonghua Yi Xue Za Zhi. 2005 85 : 1831 5.
70. Li X, Meng Y, Yang XS, Wu PS, Zhang ZS. Aldosterone stimulating PDGF-B expression in HSC via activation of EGR-1. Zhonghua Gan Zang Bing Za Zhi. 2005 13 : 567 70.
71. Saxena NK, Ikeda K, Rockey DC, Friedman SL, Anania FA. Leptin in hepatic fibrosis: evidence for increased collagen production in stellate cells and lean littermates of ob/ob mice. Hepatology. 2002 35 : 762 71.
72. Margis R, Pinheiro-Margis M, da Silva LC, Borojevic R. Effects of retinol on proliferation, cell adherence and extracellular matrix synthesis in a liver myofibroblast or lipocyte cell line (GRX). Int J Exp Pathol. 1992 73 : 125 35.
73. Guma FCR, Mello TG, Mermelstein CS, Fortuna VA, Wofchuk ST, Gottfried C, Guaragna RM, Costa ML, Borojevic R. Intermediate filaments modulation in an in vitro model of the hepatic stellate cell activation or conversion into the lipocyte phenotype. Biochem Cell Biol. 2001 79 : 409 17.
74. Zhao C, Chen W, Yang L, Chen L, Stimpson SA, Diehl AM. PPAR_ agonists prevent TGF_1/Smad3- signaling in human hepatic stellate cells. Biochem Biophys Res Commun. 2006 350 : 385 91.
75. Shi YF, Zhang Q, Cheung PY, Shi L, Fong CC, Zhang Y, Tzang CH, Chan BP, Fong WF, Chun J, Kung HF, Yang M. Effects of rhDecorin on TGF-_1 induced human hepatic stellate cells LX-2 activation. Biochim Biophys Acta. 2006 1760 : 1587 95.
76. Cao Q, Mak KM, Ren C, Lieber CS. Leptin stimulates tissue inhibitor of metalloproteinase-1 in human hepatic stellate cells: respective roles of the JAK/STAT and JAK-mediated H2 O2-dependant MAPK pathways. J Biol Chem. 2004 279 : 4292 304.
77. Cao Q, Mak KM, Lieber CS. Leptin enhances _1(I) collagen gene expression in LX-2 human hepatic stellate cells through JAK-mediated H2 O2-dependent MAPK pathways. J Cell Biochem. 2006 97 : 188 97.
78. Cao Q, Mak KM, Lieber CS. Leptin represses matrix metalloproteinase-1 gene expression in LX2 human hepatic stellate cells. J Hepatol. 2007 46 : 124 33.
79. Hui AY, Cheng AS, Chan HL, Go MY, Chan FK, Sakata R, Ueno T, Sata M, Sung JJ. Effect of prostaglandin E2 and prostaglandin I2 on PDGFinduced proliferation of LI90, a human hepatic stellate cell line. Prostaglandins Leukot Essent Fatty Acids. 2004 71 : 329 33.
80. Ueki N, Taguchi T, Takahashi M, Adachi M, Ohkawa T, Amuro Y, Hada T, Higashino K. Inhibition of hyaluronan synthesis by vesnarinone in cultured human myofibroblasts. Biochim Biophys Acta. 2000 1495 : 160 7.
81. Sugimoto R, Enjoji M, Nakamuta M, Ohta S, Kohjima M, Fukushima M, Kuniyoshi M, Arimura E, Morizono S, Kotoh K, Nawata H. Effect of IL-4 and IL-13 on collagen production in cultured LI90 human hepatic stellate cells. Liver Int. 2005 25 : 420 8.
82. Kanno K, Tazuma S, Nishioka T, Hyogo H, Chayama K. Angiotensin II participates in hepatic inflammation and fibrosis through MCP-1 expression. Dig Dis Sci. 2005 50 : 942 8.
83. Migita K, Maeda Y, Abiru S, Nakamura M, Komori A, Yokoyama T, Takii Y, Mori T, Yatsuhashi H, Eguchi K, Ishibashi H. Immunosuppressant FK506 inhibits matrix metalloproteinase-9 induction in TNF- _-stimulated human hepatic stellate cells. Life Sci. 2006 78 : 2510 5.
84. Ozaki I, Zhao G, Mizuta T, Ogawa Y, Hara T, Kajihara S, Hisatomi A, Sakai T, Yamamoto K. Hepatocyte growth factor induces collagenase (matrix metalloproteinase-1) viathe transcription factor Ets-1 in human hepatic stellate cell line. J Hepatol. 2002 36 : 169 78.