Advertisement

Isolating and Culturing Mouse Podocyte Cells to Study Diabetic Nephropathy

  • Elisavet VasilopoulouEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2067)

Abstract

Diabetic nephropathy is associated with injury and loss of podocytes, specialized epithelial cells that are critical for glomerular filtration. This chapter describes a method of isolating and culturing podocyte cells from mouse adult kidneys. In this way, podocytes with genetic modifications can be obtained from transgenic animals and they can be used to study the effects of the diabetic environment in vitro.

Key words

Diabetes Diabetic nephropathy Glomerulus Kidney Podocyte 

Notes

Acknowledgment

This work was supported by a Kidney Research UK Post-Doctoral Fellowship (PDF8/2015) to EV.

References

  1. 1.
    Alpers CE, Hudkins KL (2011) Mouse models of diabetic nephropathy. Curr Opin Nephrol Hypertens 20:278–284CrossRefGoogle Scholar
  2. 2.
    Gnudi L (2012) Cellular and molecular mechanisms of diabetic glomerulopathy. Nephrol Dial Transplant 27:2642–2649CrossRefGoogle Scholar
  3. 3.
    Gnudi L, Benedetti S, Woolf AS, Long DA (2015) Vascular growth factors play critical roles in kidney glomeruli. Clin Sci (Lond) 129:1225–1236CrossRefGoogle Scholar
  4. 4.
    Lim AK, Tesch GH (2012) Inflammation in diabetic nephropathy. Mediat Inflamm 2012:146154CrossRefGoogle Scholar
  5. 5.
    Tesch GH (2008) MCP-1/CCL2: a new diagnostic marker and therapeutic target for progressive renal injury in diabetic nephropathy. Am J Physiol 294:F697–F701Google Scholar
  6. 6.
    Saleem MA, O'Hare MJ, Reiser J, Coward RJ, Inward CD, Farren T, Xing CY, Ni L, Mathieson PW, Mundel P (2002) A conditionally immortalized human podocyte cell line demonstrating nephrin and podocin expression. J Am Soc Nephrol 13:630–638PubMedGoogle Scholar
  7. 7.
    Mundel P, Reiser J, Zuniga Mejia Borja A, Pavenstadt H, Davidson GR, Kriz W, Zeller R (1997) Rearrangements of the cytoskeleton and cell contacts induce process formation during differentiation of conditionally immortalized mouse podocyte cell lines. Exp Cell Res 236:248–258CrossRefGoogle Scholar
  8. 8.
    Lay AC, Hurcombe JA, Betin VMS, Barrington F, Rollason R, Ni L, Gillam L, Pearson GME, Østergaard MV, Hamidi H, Lennon R, Welsh GI, Coward RJM (2017) Prolonged exposure of mouse and human podocytes to insulin induces insulin resistance through lysosomal and proteasomal degradation of the insulin receptor. Diabetologia 60(11):2299–2311.  https://doi.org/10.1007/s00125-017-4394-0CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Kim Y, Lim JH, Kim MY, Kim EN, Yoon HE, Shin SJ, Choi BS, Kim Y-S, Chang YS, Park CW (2018) The adiponectin receptor agonist adiporon ameliorates diabetic nephropathy in a model of type 2 diabetes. J Am Soc Nephrol 29(4):1108–1127CrossRefGoogle Scholar
  10. 10.
    Qi W, Keenan HA, Li Q, Ishikado A, Kannt A, Sadowski T, Yorek MA, Wu I-H, Lockhart S, Coppey LJ, Pfenninger A, Liew CW, Qiang G, Burkart AM, Hastings S, Pober D, Cahill C, Niewczas MA, Israelsen WJ, Tinsley L, Stillman IE, Amenta PS, Feener EP, Vander Heiden MG, Stanton RC, King GL (2017) Pyruvate kinase M2 activation may protect against the progression of diabetic glomerular pathology and mitochondrial dysfunction. Nat Med 23(6):753–762CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Medway School of PharmacyUniversity of KentChathamUK
  2. 2.Developmental Biology and CancerUCL GOS Institute of Child HealthLondonUK

Personalised recommendations