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Cyst Swelling Assay for ADPKD

Polycystic Kidney Disease

Murine and human models available

Crown Bioscience offers a unique high content screening platform for Autosomal Dominant Polycystic Kidney Disease (ADPKD). A 384 well plate 3D assay format enables the recapitulation of the in vivo disease phenotype with robust functional readouts.

cb_cyst-in-3d-culture-01Murine cell line to model ADPKD

  • Murine Pkd1-/- collecting duct cells develop into cysts in 3D culture.
  • cAMP- dependent cyst swelling is induced by Forskolin or Prostaglandin E2.
  • Test compounds are added and the effect on cyst swelling is determined.
  • This robust model is suited to high throughput screening and also enables detailed phenotypic analysis of treatment effect and the discrimination between efficacy and toxicity.

ADPKD Patient derived ex vivo cultures

This ex vivo assay uses material collected from nephrectomy tissue from PKD patients. These tissues form cysts when cultured in a 3D environment. The cultures are bio- banked at low passage for subsequent compound testing. Cyst swelling can be driven by the addition of agonists such as desmopressin or forskolin.


Information about mutation status of these tissues is available

cb_robust-functional-readout-01Robust functional readout

Pkd1 -/- mouse collecting duct cells were cultured in 3D. After 4 days, cultures were exposed to Forskolin and reference compounds for 72h. Their capability to inhibit the cyst swelling were compared to Solvent control (0.2% DMSO) and Stimulant only control (Forskolin).





Cells that were harvested from ADPKD patients, were seeded in 384 well plates in 3D. After 24h, cultures were exposed to stimulant with reference compounds for 48h. Left: forskolin (green) or desmopressin (orange) in combination with tolvaptan (data not normalized). Right: Reference compounds were compared to positive control (Solvent only) and negative control (Desmopressin (ddAVP) only or Forskolin only) for their capability to inhibit the cyst swelling. Data was normalized to Solvent only (0%) and Desmopressin (ddAVP) only (100%)

cb-cyst-swelling-toxicity-01Discriminating between inhibition of cyst swelling and toxicity

Analysis of multiple morphological features associated with inhibition of cyst swelling and cyto- and nuclear toxicity enables the discrimination of compounds with high therapeutic potential (yellow arrow) and those which are associated with adverse toxic responses (red arrow)

Assay Principle

Key Advantages

  • High content phenotypic evaluation by 3D imaging
  • Sensitive and robust functional read-outs
  • Separation of toxic from effective compounds
  • Optimal in vitro to in vivo translation
  • Easily up-scalable assay


cb_3d-screening-platform-01After pre-culture, cells are seeded in an extracellular matrix in 384 well plates where they spontaneously form cysts. This typically takes 1-4 days, and is followed by stimulation of cyst swelling by addition of a swelling inducing compound such as vasopressin or forskolin. Simultaneously treatment that aims at inhibiting the swelling is added for 48-72h.

Set-up can be adapted to accommodate different research questions


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