title: Ex vivo analysis of renal proximal tubular cells.
abstract: BACKGROUND
  Experimental models are inevitably a compromise between accurately reproducing a pathological situation and schematically simplifying it, which is intended to provide both relevance and conclusiveness. In-vivo models are very relevant, but multiple cell-types undergoing various changes may hinder the observation of individual molecular events.
  
  
  RESULTS
  Here, we describe a method for analyzing and isolating specific cell types from the kidney and studying the phenotype they have acquired in vivo. Using flow cytometry, immunofluorescence, and RT-PCR, we show that our method is suitable for studying and isolating proximal tubular cells with an anti Prominin-1 antibody. Kidneys are subjected to mechanical dissociation followed by flow-cytometry analysis. Hundreds of thousands of proximal tubular cells are then isolated by magnetic separation followed by direct analysis or primary cell culture. Using our method, we detect phenotypic changes in the proximal tubular cells after renal ischemia reperfusion, and we isolate the proximal tubular cells, with a purity over 80%.
  
  
  CONCLUSIONS
  This method is efficient, quick, simple, and cheap, and should be useful for studying cell-type specific parameters after in vivo experimental studies. It is also a simple method to obtain a specific primary cell culture from any animal strain.
authors.affiliation: INSERM, UMR-S1155, Paris, France. davidlegouis@gmail.com.
authors.forename: David
authors.initials: D
authors.lastname: Legouis
authors.affiliation: INSERM, UMR-S1155, Paris, France. aurelbataille@gmail.com.
authors.forename: Aurélien
authors.initials: A
authors.lastname: Bataille
authors.affiliation: INSERM, UMR-S1155, Paris, France. alexandre.hertig@tnn.aphp.fr.|UPMC, Sorbonne Universités, Paris, 6, France. alexandre.hertig@tnn.aphp.fr.|Urgences Néphrologiques et Transplantation Rénale, APHP, Paris, France. alexandre.hertig@tnn.aphp.fr.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Hertig
authors.affiliation: INSERM, UMR-S1155, Paris, France. sophie.vandermeersch@upmc.fr.
authors.forename: Sophie
authors.initials: S
authors.lastname: Vandermeersch
authors.affiliation: INSERM, UMR-S1155, Paris, France. noemie.simon.tillaux@gmail.com.
authors.forename: Noémie
authors.initials: N
authors.lastname: Simon
authors.affiliation: INSERM, UMR-S1155, Paris, France. eric.rondeau@tnn.aphp.fr.|UPMC, Sorbonne Universités, Paris, 6, France. eric.rondeau@tnn.aphp.fr.|Urgences Néphrologiques et Transplantation Rénale, APHP, Paris, France. eric.rondeau@tnn.aphp.fr.
authors.forename: Eric
authors.initials: E
authors.lastname: Rondeau
authors.affiliation: INSERM, UMR-S1155, Paris, France. galichon@orange.fr.|UPMC, Sorbonne Universités, Paris, 6, France. galichon@orange.fr.|Urgences Néphrologiques et Transplantation Rénale, APHP, Paris, France. galichon@orange.fr.
authors.forename: Pierre
authors.initials: P
authors.lastname: Galichon
mesh_terms: D000071916:AC133 Antigen; D000818:Animals; D000906:Antibodies / Q000276:immunology; D015703:Antigens, CD / Q000276:immunology / Q000378:metabolism; D002469:Cell Separation; D002478:Cells, Cultured; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000276:immunology / Q000378:metabolism; D018960:Hyaluronan Receptors / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000166:cytology*; D051379:Mice; D008810:Mice, Inbred C57BL; D010455:Peptides / Q000276:immunology / Q000378:metabolism; D010641:Phenotype
chemical_list: D000071916:AC133 Antigen; D000906:Antibodies; D015703:Antigens, CD; D006023:Glycoproteins; D018960:Hyaluronan Receptors; D010455:Peptides; C000606024:Prom1 protein, mouse