title: Isolation and Primary Culture of Murine Podocytes with Proven Origin.
abstract: Genetic studies on hereditary kidney diseases and in vivo experimental model studies have revealed a critical role for the podocyte in glomerular function and disease. Primary podocyte cultures as well as immortalized podocyte cell lines have been used extensively to study podocyte function. Although, primary cells often more closely resemble the in vivo cells, they may have only a finite replicative life span before they reach senescence. Therefore, the success of studies using primary cell cultures depends on standardized isolation and culture protocols that allow reproducible generation of stable primary cultures.This chapter describes the isolation of primary podocytes with a proven origin using the novel technology of cell-specific genetic tagging. Podocytes are isolated from glomeruli from a podocyte-specific transgenic reporter mouse. The podocyte-specific reporter gene beta-galactosidase is used to identify and specifically isolate the labeled podocytes from other glomerular cells by FACS.
authors.affiliation: Department of Internal Medicine II, Nephrology and Clinical Immunology, RWTH Aachen University Hospital, Pauwelsstrasse 30, 52074, Aachen, Germany.|Department of Pathology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors.forename: Bart
authors.initials: B
authors.lastname: Smeets
authors.affiliation: Department of Internal Medicine II, Nephrology and Clinical Immunology, RWTH Aachen University Hospital, Pauwelsstrasse 30, 52074, Aachen, Germany.
authors.forename: Nazanin
authors.initials: N
authors.lastname: Kabgani
authors.affiliation: Department of Internal Medicine II, Nephrology and Clinical Immunology, RWTH Aachen University Hospital, Pauwelsstrasse 30, 52074, Aachen, Germany. mmoeller@ukaachen.de.
authors.forename: Marcus J
authors.initials: MJ
authors.lastname: Moeller
mesh_terms: D000818:Animals; D002469:Cell Separation / Q000379:methods; D051379:Mice; D050199:Podocytes / Q000166:cytology; D061251:Primary Cell Culture / Q000379:methods
keywords: Cell culture; Cell-specific tagging; Glomerulus; Pod-rtTA/LC1/R26R mice; Podocytes; Primary cell culture