Ex vivo analysis of renal proximal tubular cells

BMC Cell Biology

Pierre Galichon¹  Eric Rondeau¹  Noémie Simon²  Sophie Vandermeersch²  Alexandre Hertig¹  Aurélien Bataille²  David Legouis² 

[1] Urgences Néphrologiques et Transplantation Rénale, APHP, Paris, France;INSERM, UMR-S1155, Paris, France

关键词: In vitro;    In vivo;    Primary cell cultures;    Cell phenotype;    Experimental models;    Organ physiology;   

Others  :  1150964
DOI  :  10.1186/s12860-015-0058-4

 received in 2014-10-23, accepted in 2015-03-02,  发布年份 2015

【 摘 要 】

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.

【 授权许可】

   
2015 Legouis et al.; licensee BioMed Central.

【 图 表 】

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