Nonnis Simona, Maffioli Elisa, Zanotti Lucia, Santagata Fabiana, Negri Armando, Viola Antonella, Elliman Stephen, Tedeschi Gabriella
Dipartimento di Scienze Veterinarie e Sanità Pubblica, Università degli Studi di Milano, Via Celoria, 10, 20133 Milan, Italy.
Fondazione Filarete, Viale Ortles, 22/4, 20139 Milan, Italy.
EuPA Open Proteom. 2016 Jan 7;10:28-30. doi: 10.1016/j.euprot.2016.01.005. eCollection 2016 Mar.
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 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.
间充质基质细胞分泌组的分析对于识别涉及这些细胞的过程中的关键参与者至关重要。由于在基于质谱的条件培养基(CM)分析中,添加了胎牛血清(FBS)的蛋白质,真正分泌的蛋白质可能难以检测。我们比较了不同的生长条件,以确定不同FBS浓度对污染牛蛋白质的真正分泌的人类蛋白质数量和含量的影响。结果表明,为了尽量减少干扰,细胞应在FBS存在下生长至汇合,然后在收集分泌组之前转移到无血清培养基中。