Glomeruli Isolation: A Technique to Obtain Intact Glomeruli from Murine Kidney

Abstract

Source: Eymael, J. et al. Glomerular Outgrowth as an Ex Vivo Assay to Analyze Pathways Involved in Parietal Epithelial Cell Activation. J. Vis. Exp. (2020)

This video describes the method to isolate intact glomeruli from mouse kidneys. The encapsulated and decapsulated glomeruli thus obtained can further be used for culturing parietal epithelial cell outgrowths from the murine kidney.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

NOTE: Untreated, healthy wild-type (WT) mice (n = 4) and cd44-/- (n = 4) mice were sacrificed at the age of 12−16 weeks. Both male and female mice were used. All mice were on the C57Bl/6 background.

1. Mouse Kidney Dissection

1. Sacrifice healthy WT mice or genetically altered mice by cervical dislocation.
2. Dissect whole mouse kidneys directly after sacrificing the mice. For this, perform a medi....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
24-well cell culture plate	Corning Costar
Digital inverted light microscope	Westburg, EVOS fl microscope
Dulbecco's Modified Eagle's medium	Lonza
Fetal Bovine Serum	Lonza
Hanks' Balanced Salt Solution	Gibco
Petri dish	Sarstedt		Size 100
Scalpel	Dahlhausen		Size 10
Sieves	Endecotts Ltd		Size: 300 µm, 75 µm, 53 µm, steel
Ultra-Low Attachment Microplates	Corning Costar		6-well plates
Syringe	BD Plastipak		Size: 20 mL