Abstract
Renal physiology and pathology are complex systems that are best studied in whole living organisms. This, however, is often restricted by our desire to limit the number of animal experiments undertaken and to replace them with relevant in vitro models that can be used as surrogates for the system under test. Primary culture cells are derived directly from the relevant tissue and therefore correlate more closely with the system under examination. Although the tissue of origin is not always readily available for culture and cells may quickly change their phenotype after only a few passages, they can be used in many circumstances to validate results obtained from closely related cell lines and to confirm vital protein expression patterns. This chapter outlines methods by which proximal tubular epithelial cells and renal interstitial fibroblasts can be isolated and characterized from human renal nephrectomy tissue.
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References
Racusen, LC, Monteil, C, Sgrignoli, A, Lucskay, M, Marouillat, S, Rhim, JG & Morin, JP: Cell lines with extended in vitro growth potential from human renal proximal tubule: characterization, response to inducers, and comparison with established cell lines. J Lab Clin Med 129:318–29, 1997.
Ryan, MJ, Johnson, G, Kirk, J, Fuerstenberg, SM, Zager, RA & Torok-Storb, B: HK-2: an immortalized proximal tubule epithelial cell line from normal adult human kidney. Kidney Int 45:48–57, 1994.
Muller, GA & Rodemann, HP: Characterization of human renal fibroblasts in health and disease: I. Immunophenotyping of cultured tubular epithelial cells and fibroblasts derived from kidneys with histologically proven interstitial fibrosis. Am J Kidney Dis 17:680–3, 1991.
Rodemann, HP & Muller, GA: Characterization of human renal fibroblasts in health and disease: II. In vitro growth, differentiation, and collagen synthesis of fibroblasts from kidneys with interstitial fibrosis. Am J Kidney Dis 17:684–6, 1991.
Grimwood, L & Masterson, R: Propagation and culture of renal fibroblasts. Methods Mol Biol 466:25–37, 2009.
Nachlas, MM, Monis, B, Rosenbatt, D & Seligman, AM: Improvement in the histochemical localization of leucine aminopeptidase with a new substrate, L-leucyl-4-methoxy-2-naphthylamide. J Biophys Biochem Cytol 7:261–4, 1960.
Rutenburg, AM, Kim, H, Fischbein, JW, Hanker, JS, Wasserkrug, HL & Seligman, AM: Histochemical and ultrastructural demonstration of gamma-glutamyl transpeptidase activity. J Histochem Cytochem 17:517–26, 1969.
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Sharpe, C.C., Dockrell, M.E.C. (2012). Primary Culture of Human Renal Proximal Tubule Epithelial Cells and Interstitial Fibroblasts. In: Mitry, R., Hughes, R. (eds) Human Cell Culture Protocols. Methods in Molecular Biology, vol 806. Humana Press. https://doi.org/10.1007/978-1-61779-367-7_12
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DOI: https://doi.org/10.1007/978-1-61779-367-7_12
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