Vibrodissociation method for isolation of defined nephron segments from human and rodent kidneys. Am J Physiol Renal Physiol 2019 Nov 01;317(5):F1398-F1403
Date
10/08/2019Pubmed ID
31588797Pubmed Central ID
PMC6879945DOI
10.1152/ajprenal.00448.2019Scopus ID
2-s2.0-85074674820 (requires institutional sign-in at Scopus site) 7 CitationsAbstract
Our current knowledge of the properties of renal ion channels responsible for electrolytes and cell energy homeostasis mainly relies on rodent studies. However, it has not been established yet to what extent their characteristics can be generalized to those of humans. The present study was designed to develop a standardized protocol for the isolation of well-preserved glomeruli and renal tubules from rodent and human kidneys and to assess the functional suitability of the obtained materials for physiological studies. Separation of nephron segments from human and rodent kidneys was achieved using a novel vibrodissociation technique. The integrity of isolated renal tubules and glomeruli was probed via electrophysiological analysis and fluorescence microscopy, and the purity of the collected fractions was confirmed using quantitative RT-PCR with gene markers for specific cell types. The developed approach allows rapid isolation of well-preserved renal tubules and glomeruli from human and rodent kidneys amenable for electrophysiological, Ca2+ imaging, and omics studies. Analysis of the basic electrophysiological parameters of major K+ and Na+ channels expressed in human cortical collecting ducts revealed that they exhibited similar biophysical properties as previously reported in rodent studies. Using vibrodissociation for nephron segment isolation has several advantages over existing techniques: it is less labor intensive, requires little to no enzymatic treatment, and produces large quantities of well-preserved experimental material in pure fractions. Applying this method for the separation of nephron segments from human and rodent kidneys may be a powerful tool for the indepth assessment of kidney function in health and disease.
Author List
Isaeva E, Fedoriuk M, Bohovyk R, Klemens CA, Khedr S, Golosova D, Levchenko V, El-Meanawy A, Palygin O, Staruschenko AAuthors
Ashraf El-Meanawy MD, PhD Professor in the Medicine department at Medical College of WisconsinOlena Isaeva PhD Assistant Professor in the Cell Biology, Neurobiology and Anatomy department at Medical College of Wisconsin
MESH terms used to index this publication - Major topics in bold
AnimalsCalcium
Histocytological Preparation Techniques
Humans
Mice
Nephrons
Rats
Rats, Inbred Dahl
Vibration
