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Culture of Three-Dimensional Madin–Darby Canine Kidney (MDCK) Cysts for In Vitro Drug Testing in Polycystic Kidney Disease

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Kidney Research

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2664))

Abstract

The formation and growth of kidney cysts (fluid-filled structures lined by epithelial cells) is the primary pathological abnormality in polycystic kidney disease (PKD). Multiple molecular pathways are disrupted in kidney epithelial precursor cells, which lead to altered planar cell polarity, increased proliferation, and fluid secretion, which together with extracellular matrix remodelling culminates in the formation and growth of cysts. Three-dimensional (3D) in vitro cyst models serve as suitable preclinical models to screen candidate drugs for PKD. Madin–Darby Canine Kidney (MDCK) epithelial cells form polarized monolayers with a fluid-filled lumen when suspended in a collagen gel, and their growth is accelerated with the addition of forskolin, a cyclic adenosine monophosphate (cAMP) agonist. Candidate drugs for PKD can be screened for their ability to modulate growth of forskolin-treated MDCK cysts by measuring and quantifying cyst images acquired at progressive timepoints. In this chapter, we describe the detailed methods for the culture and growth of MDCK cysts in a collagen matrix and a protocol for their use in testing candidate drugs to prevent cyst formation and growth.

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Author information

Authors and Affiliations

  1. Michael Stern Laboratory for Polycystic Kidney Disease, Westmead Institute for Medical Research, The University of Sydney, Westmead, NSW, Australia

    Sayanthooran Saravanabavan & Gopala K. Rangan

  2. Department of Renal Medicine, Westmead Hospital, Western Sydney Local Health District, Westmead, NSW, Australia

    Sayanthooran Saravanabavan & Gopala K. Rangan

  3. Westmead Clinical School, Faculty of Medicine and Health, The University of Sydney, Westmead, NSW, Australia

    Gopala K. Rangan

Authors
  1. Sayanthooran Saravanabavan
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  2. Gopala K. Rangan
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Corresponding author

Correspondence to Gopala K. Rangan .

Editor information

Editors and Affiliations

  1. Department of Nephrology, The Royal Melbourne Hospital, Parkville, Melbourne, VIC, Australia

    Tim D. Hewitson

  2. Department of Nephrology, The Royal Melbourne Hospital, Parkville, Melbourne, VIC, Australia

    Nigel D. Toussaint

  3. Department of Nephrology, The Royal Melbourne Hospital, Parkville, Melbourne, VIC, Australia

    Edward R. Smith

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Saravanabavan, S., Rangan, G.K. (2023). Culture of Three-Dimensional Madin–Darby Canine Kidney (MDCK) Cysts for In Vitro Drug Testing in Polycystic Kidney Disease. In: Hewitson, T.D., Toussaint, N.D., Smith, E.R. (eds) Kidney Research. Methods in Molecular Biology, vol 2664. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3179-9_10

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  • DOI: https://doi.org/10.1007/978-1-0716-3179-9_10

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3178-2

  • Online ISBN: 978-1-0716-3179-9

  • eBook Packages: Springer Protocols

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