Kidney Research Experimental Protocols

This third edition provides updated and new chapters on recent innovations in basic renal research. Chapters are divided into three sections covering in vitro, in vivo, and ex vivo models of kidney disease, recent advances in imaging techniques and protocols for performing analytical and functional measurements in the kidney; techniques that are both topical and of widespread relevance to the study of kidney biology and disease. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and methods, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.

Authoritative and cutting-edge, Kidney Research: Experimental Protocols, Third Edition aims to be comprehensive guide for researchers in the field.

Restoration of podocyte phenotype in culture.- Protocols for the propagation, culture and characterization of kidney fibroblasts.- Isolation of rat glomeruli and propagation of mesangial cells to study the kidney in health and disease.- In vitro models to evaluate molecular permeability of the kidney filtration barrier.- Generating human glomeruli from pluripotent stem cells for disease modelling and drug screening.- Large-scale production of kidney organoids from human pluripotent stem cells.- Construction of a multitubular perfusable kidney-on-chip for the study of renal disease.- A “kidney-on-the-chip” model composed of primary human tubular, endothelial, and white blood cells.- Standardized protocol for the preparation of precision-cut kidney slices:  a translational model of renal fibrosis.- Culture of three-dimensional Madin-Darby Canine Kidney (MDCK) cysts for in vitro drug testing in polycystic kidney disease.- Zebrafishmodel to study podocyte function within the glomerular filtration barrier.- Magnetic Resonance Imaging (MRI) analysis of tissue sodium concentration in chronic kidney disease.- Determination of interstitial collagen deposition and related topography using the second harmonic generation-based Histoindex platform.- Quantitative imaging of podocyte foot processes in the kidney using confocal and STED microscopy.- Synthesis and expression of a targeted, ferritin-based tracer for PET imaging of kidney glomeruli.- Multiplex in situ hybridization in the study of acute kidney injury.- Spatial transcriptomics in kidney tissue.- Assessment of kidney mitochondrial function by high-resolution respirometry, transmission electron microscopy and histological techniques.- Transdermal measurement of glomerular filtration rate in preclinical research.- The BioHybrid Assay – a novel method for determining calcification propensity.- Quantification of calciprotein particles (CPPs) in serum/plasma samples using a fluorescent bisphosphonate.- Monitoring of Rho GTPase Activity in Podocytes.