Isolation and cultivation of myofibroblasts from rats liver using explantation method

Abstract

During liver fibrosis development connective tissueis produced by myofibroblasts that could originate fromtwo hepatic populations: hepatic stellate cells and portalfibroblasts. A marker of myofibroblasts is the expressionof ƒ-smooth muscle actin (ƒ-SMA). Distinctive feature ofmyofibroblasts, derived from hepatic stellate cells, is thepreservation of the hepatic stellate cells marker expression -desmin. The processes of activation, proliferation and cellstrans-differentiation into myofibroblasts are closely relatedto the activity of transcription factor NF-kB and its inhibitorIkBƒ. The aim of our work was to obtain a culture of hepaticmyofibrobasts, to study their origin, phenotype, relationsbetween NF-kB and IkBƒ expression and the processes ofactivation and cells trans-differentiation into myofibroblasts.For this purpose we isolated heterogeneous population ofcells from rat liver by the method of explantation. Almostall the cells had desmin and ƒ-SMA expression. On thisbasis, we suppose that these myofibroblasts were hepaticstellate cells derivatives, and singular desmin-negative cellsoriginated from portal fibroblasts. Thus, hepatic stellate cellshave major potential to activation, growth, proliferation andtransdifferentiation into myofibroblasts in comparison toportal fibroblasts. Activated state of the cells was confirmedby stable expression of NF-kB and its inhibitor IkBƒ in all thecells throughout the whole experiment.

Keywords

hepatic stellate cells / portal fibroblasts / transcription factor NF-kB / desmin / ƒ-SMA / inhibitor IkBƒ

References

[1] Gumerova A., Abdulkhakov S. Cell sources of liver development and regeneration. Saarbrucken: LAP LAMBERT Academic Publishing GmbH &Co.KG; 2012.

[2] Гумерова А.А. Киясов А.П. Могут ли перисинусоидальные клетки быть региональными стволовыми [прогениторными] клет- ками печени? Клеточная трансплантология и тканевая инженерия 2010; 5(1): 33-40.

[3] Ramadori G., Saile B. Mesenchymal cells in the liver - one cell type or two? Liver 2002; 22(4): 283-94.

[4] Iwaisako K., Brenner D.A., Kisseleva T. What's new in liver fibrosis? The origin of myofibroblasts in liver fibrosis. J. Gastroenterol. Hepatol. 2012; 27(Suppl 2): 65-8.

[5] Eng F.J., Friedman S.L. Transcriptional regulation in hepatic stellate cells. Semin. Liver Dis. 2001; 21(3): 385-95.

[6] Kisseleva T., Brenner D.A. Anti-fibrogenic strategies and the regression of fibrosis. Best Pract Res Clin Gastroenterol. 2011; 25(2): 305-17.

[7] Knook D.L., Seffelaar A.M., de Leeuw A.M. Fat-storing cells of the rat liver. Their isolation and purification. Exp. Cell Res. 1982; 139(2): 468-71.

[8] Bosselut N., Housset C., Marcelo P. et al. Distinct proteomic features of two fibrogenic liver cell populations: hepatic stellate cells and portal myofibroblasts. Proteomics 2010; 10(5): 1017-28.

[9] Knittel T., Kobold D., Saile B. et al. Rat liver myofibroblasts and hepatic stellate cells: different cell populations of the fibroblast lineage with fibrogenic potential. Gastroenterology 1999; 117(5): 1205-21.

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