Autosomal Dominant Polycystic Kidney Disease (ADPKD) is a slowly progressive genetic renal disorder that is caused by mutations in either the PKD1 gene (polycystin 1) or the PKD2 gene (polycystin 2). ADPKD is characterized by the formation of fluid filled cysts in both kidneys leading eventually to end-stage renal disease. Several in vivo and in vitro models of ADPKD exist, although many of them fall short of fully recapitulating the human disease phenotype, driving the failure of most therapeutic candidates in human clinical trials.
Development of the 3D ADPKD Model System
Human ADPKD kidneys are ethically sourced from our tissue exchange partners where we convert raw tissue into useable cells that are able to form cysts in 3D Biogel cultures. These cells become the cornerstone of our unique 3D in vitro system.
Our Model System
We have developed a unique 3D Biogel-based platform in 384-well tissue culture format utilizing cells isolated from individual cysts on human ADPKD donor kidneys. Cultures from each individual cyst are genotyped to determine the PKD1 or PKD2 mutation. Interestingly, from 9 human ADPKD donor tissues to date (8 with PKD1 mutations, 1 with a PKD2 mutation), we have discovered that only a subset of donors are prone to second somatic hits in single cyst-derived cultures, in addition to the germline mutation. These cells form cysts over days to weeks in culture that can be tracked by high-content imaging with cyst size and number being quantified through algorithm-based image analysis. This platform can be used for screening or validation of candidate drugs and provides significant advancement in throughput and pathophysiologic relevancy.
Industry Standards Sample Data Set
Human ADPKD cells were exposed to either DMSO vehicle or treatment over 13 days in 3D Culture. Live-cell images and CellTiterGlo (CTG) were captured at endpoint. Representative images were selected from a concentration (0.1µM) where no cytotoxicity was seen between any therapeutic. Images were analyzed for Cyst Number and Size by proprietary image analysis software. Note that no stimulus such as forskolin or arginine-vasopressin was used in this experiment, although can be added to accelerate cyst expansion.
Our advanced 3D live-cell 384-well based cystogenesis assay, utilizing human ADPKD cells, provides enough throughput for screening with enough pathophysiologic relevancy to help validate therapeutic efficacy thus providing crucial information for key decision points along your therapeutics’ developmental pathway.
Please inquire to learn more about experimental modalities, conditions, and customizable options – including the ability to isolate RNA or protein for further downstream analysis.