Primary cultures of glomerular parietal epithelial cells or podocytes with proven origin.

Parietal epithelial cells (PECs) are crucially involved in the pathogenesis of rapidly progressive glomerulonephritis (RPGN) as well as in focal and segmental glomerulosclerosis (FSGS). In this study, transgenic mouse lines were used to isolate pure, genetically tagged primary cultures of PECs or po...

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Autores principales:	Nazanin Kabgani, Tamara Grigoleit, Kevin Schulte, Antonio Sechi, Sibille Sauer-Lehnen, Carmen Tag, Peter Boor, Christoph Kuppe, Gregor Warsow, Sandra Schordan, Jörg Mostertz, Ravi Kumar Chilukoti, Georg Homuth, Nicole Endlich, Frank Tacke, Ralf Weiskirchen, Georg Fuellen, Karlhans Endlich, Jürgen Floege, Bart Smeets, Marcus J Moeller
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b3ba4fc1cb2d4a5d9ce95846f3e4eec8
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spelling	oai:doaj.org-article:b3ba4fc1cb2d4a5d9ce95846f3e4eec82021-11-18T07:21:49ZPrimary cultures of glomerular parietal epithelial cells or podocytes with proven origin.1932-620310.1371/journal.pone.0034907https://doaj.org/article/b3ba4fc1cb2d4a5d9ce95846f3e4eec82012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22529955/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Parietal epithelial cells (PECs) are crucially involved in the pathogenesis of rapidly progressive glomerulonephritis (RPGN) as well as in focal and segmental glomerulosclerosis (FSGS). In this study, transgenic mouse lines were used to isolate pure, genetically tagged primary cultures of PECs or podocytes using FACsorting. By this approach, the morphology of primary glomerular epithelial cells in culture could be resolved: Primary podocytes formed either large cells with intracytoplasmatic extensions or smaller spindle shaped cells, depending on specific culture conditions. Primary PECs were small and exhibited a spindle-shaped or polygonal morphology. In the very early phases of primary culture, rapid changes in gene expression (e.g. of WT-1 and Pax-2) were observed. However, after prolonged culture primary PECs and podocytes still segregated clearly in a transcriptome analysis--demonstrating that the origin of primary cell cultures is important. Of the classical markers, synaptopodin and podoplanin expression were differentially regulated the most in primary PEC and podocyte cultures. However, no expression of any endogenous gene allowed to differentiate between the two cell types in culture. Finally, we show that the transcription factor WT1 is also expressed by PECs. In summary, genetic tagging of PECs and podocytes is a novel and necessary tool to derive pure primary cultures with proven origin. These cultures will be a powerful tool for the emerging field of parietal epithelial cell biology.Nazanin KabganiTamara GrigoleitKevin SchulteAntonio SechiSibille Sauer-LehnenCarmen TagPeter BoorChristoph KuppeGregor WarsowSandra SchordanJörg MostertzRavi Kumar ChilukotiGeorg HomuthNicole EndlichFrank TackeRalf WeiskirchenGeorg FuellenKarlhans EndlichJürgen FloegeBart SmeetsMarcus J MoellerPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 4, p e34907 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Nazanin Kabgani Tamara Grigoleit Kevin Schulte Antonio Sechi Sibille Sauer-Lehnen Carmen Tag Peter Boor Christoph Kuppe Gregor Warsow Sandra Schordan Jörg Mostertz Ravi Kumar Chilukoti Georg Homuth Nicole Endlich Frank Tacke Ralf Weiskirchen Georg Fuellen Karlhans Endlich Jürgen Floege Bart Smeets Marcus J Moeller Primary cultures of glomerular parietal epithelial cells or podocytes with proven origin.
description	Parietal epithelial cells (PECs) are crucially involved in the pathogenesis of rapidly progressive glomerulonephritis (RPGN) as well as in focal and segmental glomerulosclerosis (FSGS). In this study, transgenic mouse lines were used to isolate pure, genetically tagged primary cultures of PECs or podocytes using FACsorting. By this approach, the morphology of primary glomerular epithelial cells in culture could be resolved: Primary podocytes formed either large cells with intracytoplasmatic extensions or smaller spindle shaped cells, depending on specific culture conditions. Primary PECs were small and exhibited a spindle-shaped or polygonal morphology. In the very early phases of primary culture, rapid changes in gene expression (e.g. of WT-1 and Pax-2) were observed. However, after prolonged culture primary PECs and podocytes still segregated clearly in a transcriptome analysis--demonstrating that the origin of primary cell cultures is important. Of the classical markers, synaptopodin and podoplanin expression were differentially regulated the most in primary PEC and podocyte cultures. However, no expression of any endogenous gene allowed to differentiate between the two cell types in culture. Finally, we show that the transcription factor WT1 is also expressed by PECs. In summary, genetic tagging of PECs and podocytes is a novel and necessary tool to derive pure primary cultures with proven origin. These cultures will be a powerful tool for the emerging field of parietal epithelial cell biology.
format	article
author	Nazanin Kabgani Tamara Grigoleit Kevin Schulte Antonio Sechi Sibille Sauer-Lehnen Carmen Tag Peter Boor Christoph Kuppe Gregor Warsow Sandra Schordan Jörg Mostertz Ravi Kumar Chilukoti Georg Homuth Nicole Endlich Frank Tacke Ralf Weiskirchen Georg Fuellen Karlhans Endlich Jürgen Floege Bart Smeets Marcus J Moeller
author_facet	Nazanin Kabgani Tamara Grigoleit Kevin Schulte Antonio Sechi Sibille Sauer-Lehnen Carmen Tag Peter Boor Christoph Kuppe Gregor Warsow Sandra Schordan Jörg Mostertz Ravi Kumar Chilukoti Georg Homuth Nicole Endlich Frank Tacke Ralf Weiskirchen Georg Fuellen Karlhans Endlich Jürgen Floege Bart Smeets Marcus J Moeller
author_sort	Nazanin Kabgani
title	Primary cultures of glomerular parietal epithelial cells or podocytes with proven origin.
title_short	Primary cultures of glomerular parietal epithelial cells or podocytes with proven origin.
title_full	Primary cultures of glomerular parietal epithelial cells or podocytes with proven origin.
title_fullStr	Primary cultures of glomerular parietal epithelial cells or podocytes with proven origin.
title_full_unstemmed	Primary cultures of glomerular parietal epithelial cells or podocytes with proven origin.
title_sort	primary cultures of glomerular parietal epithelial cells or podocytes with proven origin.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/b3ba4fc1cb2d4a5d9ce95846f3e4eec8
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Primary cultures of glomerular parietal epithelial cells or podocytes with proven origin.

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