Mouse Kidney Organoid Culture

Cystogenesis is a morphological consequence of numerous genetic diseases of the epithelium. In the kidney, the pathogenic mechanisms underlying the program of altered cell and tubule morphology are obscured by secondary effects of cyst expansion. Here, we present a 3D tubuloid system that will allow investigators to isolate the rapid changes in protein localization and gene expression that correlate with altered cell and tubule morphology during cyst initiation.

Mouse renal tubule fragments isolated from postnatal day 7 (P7) wild type mice were pulsed with a cell differentiation cocktail including glial derived neurotrophic factor (GDNF) to yield collecting duct-like tubuloid structures with appropriate polarity, primary cilium, and gene expression.

This model can be used to study early steps in cystogenesis. Mouse renal tubule fragments are isolated at P7 from PKD1flox/flox or PKD2flox/flox; Pax8rtTA; TetOCre; mTmG mice. Within hours of doxycycline-induced inactivation of PKD1 or PKD2 and loss of polycystin-1 or 2, there is significant progression from tubuloid to cyst morphology. This system allows tracking of the morphological, protein and genetic changes associated with the early phases of cystogenesis. You can find this system reported in the following publication:

GDNF drives rapid tubule morphogenesis in a novel 3D in vitro model for ADPKD
Eryn E. Dixon, Demetrios S. Maxim, Victoria L. Halperin Kuhns, Allison C. Lane-Harris, Patricia Outeda, Andrew J. Ewald, Terry J. Watnick, Paul A. Welling, Owen M. Woodward
Journal of Cell Science 2020 133: jcs249557 doi: 10.1242/jcs.249557

https://jcs.biologists.org/content/133/14/jcs249557

For more information, contact Owen M. Woodward at OWoodward@som.umaryland.edu.

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