Medical College of Wisconsin
CTSICores SearchResearch InformaticsREDCap

Improved mass spectrometric proteomic profiling of the secretome of rat vascular endothelial cells. J Proteome Res 2006 Oct;5(10):2861-4 PMID: 17022658 PMCID: PMC2814535

Abstract

Serum albumin contamination of cells cultured in vitro significantly impedes the mass spectrometric analysis of proteins secreted by the cells. Here we report a novel washing and culturing technique for rat vascular endothelial cells that considerably reduces the concentration of the commonly used additive for cell culture, bovine serum albumin (BSA), in the secretome of these cells. Cells are rinsed stringently and cultured for 24 h in serum-free media without appreciably impeding cell growth or viability. The percentage of BSA scans identified by tandem mass spectrometry (LC-MS/MS) in stringently rinsed cells (average 13.2%) was significantly lower than either the moderately rinsed or no rinse cell treatments (average 35.2% and 45.2% respectively). Furthermore, the stringent wash treatment allowed the confident identification of a larger portion of the secretome of rat endothelial cells by LC-MS/MS.

Author List

Pellitteri-Hahn MC, Warren MC, Didier DN, Winkler EL, Mirza SP, Greene AS, Olivier M

Author

Andrew S. Greene PhD Interim Vice Chair, Chief, Professor in the Biomedical Engineering department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Animals
Cattle
Cell Culture Techniques
Cells, Cultured
Culture Media
Endothelial Cells
Endothelium, Vascular
Mass Spectrometry
Proteome
Proteomics
Rats
Serum Albumin, Bovine



View this publication's entry at the Pubmed website PMID: 17022658
jenkins-FCD Prod-158 788e756cee95f97989f23472575fc754037544be