Background: Understanding podocyte biology is key to deciphering the pathogenesis of numerous glomerular diseases. However, cultivation of primary podocytes results in de-differentiation with loss of specialised architecture. Human conditionally immortalised podocytes partly overcome this problem, utilising a temperature-sensitive transgene. Conditionally immortalised murine podocytes exist, but are derived from the Immortomouse. Methods: Using retroviral temperature-sensitive SV40 transfection, we created a conditionally immortalised podocyte cell line from wild-type mice. Results: These cells develop characteristic mature podocyte morphology and robustly express slit diaphragm proteins. Functionally, these cells demonstrate comparable responses in motility and glucose uptake to human conditionally immortalised podocytes. Conclusion: Podocyte-specific transgenic mice are extensively used to study glomerular disease and this technique could be used to make podocyte cell lines from any mouse, allowing study at the cellular level. This will help characterise these disease models and add to the laboratory resources used to study podocytopathies and glomerular disease.

Keywords:
Mouse, Podocytes, Immortalisation, Slit diaphragm
1.
Abrahamson DR: Role of the podocyte (and glomerular endothelium) in building the GBM. Semin Nephrol 2012;32:342-349.
2.
Büscher AK, Weber S: Educational paper: the podocytopathies. Eur J Pediatr 2012;171:1151-1160.
3.
Jarad G, Miner JH: Update on the glomerular filtration barrier. Curr Opin Nephrol Hypertens 2009;18:226-232.
4.
Pabst R, Sterzel RB: Cell renewal of glomerular cell types in normal rats. An autoradiographic analysis. Kidney Int 1983;24:626-631.
5.
Welsh GI, Saleem MA: The podocyte cytoskeleton - key to a functioning glomerulus in health and disease. Nat Rev Nephrol 2012;8:14-21.
6.
Tryggvason K, Wartiovaara J: How does the kidney filter plasma? Physiology (Bethesda) 2005;20:96-101.
7.
Gipson DS, Massengill SF, Yao L, Nagaraj S, Smoyer WE, Mahan JD, Wigfall D, Miles P, Powell L, Lin JJ, Trachtman H, Greenbaum LA: Management of childhood onset nephrotic syndrome. Pediatrics 2009;124:747-757.
8.
Smoyer WE, Mundel P: Regulation of podocyte structure during the development of nephrotic syndrome. J Mol Med (Berl) 1998;76:172-183.
9.
Coward RJ, Saleem MA: Podocytes as a target of insulin. Curr Diabetes Rev 2011;7:22-27.
10.
Yaoita E, Yamamoto T, Takashima N, Kawasaki K, Kawachi H, Shimizu F, Kihara I: Visceral epithelial cells in rat glomerular cell culture. Eur J Cell Biol 1995;67:136-144.
11.
Saleem MA, O'Hare MJ, Reiser J, Coward RJ, Inward CD, Farren T, Xing CY, Ni L, Mathieson PW, Mundel P: A conditionally immortalized human podocyte cell line demonstrating nephrin and podocin expression. J Am Soc Nephrol 2002;13:630-638.
12.
O'Hare MJ, Bond J, Clarke C, Takeuchi Y, Atherton AJ, Berry C, Moody J, Silver AR, Davies DC, Alsop AE, Neville AM, Jat PS: Conditional immortalization of freshly isolated human mammary fibroblasts and endothelial cells. Proc Natl Acad Sci U S A 2001;98:646-651.
13.
Jat PS, Noble MD, Ataliotis P, Tanaka Y, Yannoutsos N, Larsen L, Kioussis D: Direct derivation of conditionally immortal cell lines from an H-2Kb-tsA58 transgenic mouse. Proc Natl Acad Sci U S A 1991;88:5096-5100.
14.
Shankland SJ, Pippin JW, Reiser J, Mundel P: Podocytes in culture: past, present, and future. Kidney Int 2007;72:26-36.
15.
Mundel P, Reiser J, Zúñiga Mejía Borja A, Pavenstädt H, Davidson GR, Kriz W, Zeller R: Rearrangements of the cytoskeleton and cell contacts induce process formation during differentiation of conditionally immortalized mouse podocyte cell lines. Exp Cell Res 1997;236:248-258.
16.
Ni L, Saleem M, Mathieson PW: Podocyte culture: tricks of the trade. Nephrology (Carlton) 2012;17:525-531.
17.
Satchell SC, Tasman CH, Singh A, Ni L, Geelen J, von Ruhland CJ, O'Hare MJ, Saleem MA, van den Heuvel LP, Mathieson PW: Conditionally immortalized human glomerular endothelial cells expressing fenestrations in response to VEGF. Kidney Int 2006;69:1633-1640.
18.
Sarrab RM, Lennon R, Ni L, Wherlock MD, Welsh GI, Saleem MA: Establishment of conditionally immortalized human glomerular mesangial cells in culture, with unique migratory properties. Am J Physiol Renal Physiol 2011;301:F1131-F1138.
19.
Mundel P, Heid HW, Mundel TM, Krüger M, Reiser J, Kriz W: Synaptopodin: an actin-associated protein in telencephalic dendrites and renal podocytes. J Cell Biol 1997;139:193-204.
20.
Harris JJ, McCarthy HJ, Ni L, Wherlock M, Kang H, Wetzels JF, Welsh GI, Saleem MA: Active proteases in nephrotic plasma lead to a podocin-dependent phosphorylation of VASP in podocytes via protease activated receptor-1. J Pathol 2013;229:660-671.
21.
Lee EY, Chung CH, Khoury CC, Yeo TK, Pyagay PE, Wang A, Chen S: The monocyte chemoattractant protein-1/CCR2 loop, inducible by TGF-beta, increases podocyte motility and albumin permeability. Am J Physiol Renal Physiol 2009;297:F85-F94.
22.
Herman-Edelstein M, Weinstein T, Gafter U: TGFβ1-dependent podocyte dysfunction. Curr Opin Nephrol Hypertens 2013;22:93-99.
23.
Hale LJ, Hurcombe J, Lay A, Santamaría B, Valverde AM, Saleem MA, Mathieson PW, Welsh GI, Coward RJ: Insulin directly stimulates VEGF-A production in the glomerular podocyte. Am J Physiol Renal Physiol 2013;305:F182-F188.
24.
Coward RJ, Welsh GI, Yang J, Tasman C, Lennon R, Koziell A, Satchell S, Holman GD, Kerjaschki D, Tavaré JM, Mathieson PW, Saleem MA: The human glomerular podocyte is a novel target for insulin action. Diabetes 2005;54:3095-3102.
25.
Chittiprol S, Chen P, Petrovic-Djergovic D, Eichler T, Ransom RF: Marker expression, behaviors, and responses vary in different lines of conditionally immortalized cultured podocytes. Am J Physiol Renal Physiol 2011;301:F660-F671.
26.
Roselli S, Heidet L, Sich M, Henger A, Kretzler M, Gubler MC, Antignac C: Early glomerular filtration defect and severe renal disease in podocin-deficient mice. Mol Cell Biol 2004;24:550-560.
© 2015 S. Karger AG, Basel
2015
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.