Effect of fetal bovine serum in culture media on MS analysis of mesenchymal stromal cells secretome

Q4 Biochemistry, Genetics and Molecular Biology
Simona Nonnis , Elisa Maffioli , Lucia Zanotti , Fabiana Santagata , Armando Negri , Antonella Viola , Stephen Elliman , Gabriella Tedeschi
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引用次数: 23

Abstract

The analysis of mesenchymal stromal cells secretome is fundamental to identify key players of processes involving these cells. Truly secreted proteins may be difficult to detect in MS based analysis of conditioned media (CM) due to proteins supplemented with fetal bovine serum (FBS). We compared different growth conditions to determine the effect of varying FBS concentration on the number and quantity of truly secreted human proteins vs contaminating bovine proteins. The results suggest that to minimize interference cells should be grown in presence of FBS until confluence and transferred into a serum-free medium prior to secretome collection.

Abstract Image

培养基中胎牛血清对间充质间质细胞分泌组质谱分析的影响
间充质间质细胞分泌组的分析是确定与这些细胞有关的过程的关键参与者的基础。在条件培养基(CM)的MS分析中,由于蛋白质中添加了胎牛血清(FBS),可能难以检测到真正分泌的蛋白质。我们比较了不同的生长条件,以确定不同的FBS浓度对真正分泌的人蛋白和污染的牛蛋白的数量和数量的影响。结果表明,为了减少干扰,细胞应在FBS存在下生长直到融合,并在分泌组收集之前将其转移到无血清培养基中。
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来源期刊
EuPA Open Proteomics
EuPA Open Proteomics Biochemistry, Genetics and Molecular Biology-Biochemistry
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