Variations of secretome profiles according to conditioned medium preparation: The example of human mesenchymal stem cell‐derived adipocytes

Abstract

One challenging point in analyzing cellular secretome collected as conditioned medium is cross-contamination by cell culture media components, especially bovine serum proteins. A common approach for serum removal is to wash the cells, an alternative is to grow cells using serum-free conditions. Given that the sample processing may influence the phenotype of cells and thus the secretome, it is important to establish the optimal protocol for each cell type. In this study, we compared two methods for preparing conditioned medium from human adipocytes derived from mesenchymal stem cells. Cells were either washed twice with PBS or cultured the last four days of differentiation in serum-free adipogenic medium. Gene expression of the cells was evaluated by using real-time PCR and 1D LC-MS/MS was used to compare secreted proteins present in the culture supernatants. Surprisingly, results showed significant differences in gene expression patterns of the cells and in protein content of the conditioned media and suggested that PBS washes induced severe modifications of the phenotype of cells and thus changes in protein secretion profiles. These data emphasize the significant variations in protein species related to cell manipulations and underline the importance of procedure optimization prior to any proteomic investigation.

The Comparison of Adipose Stem Cell and Placental Stem Cell in Secretion Characteristics and in Facial Antiaging

“…The phenomenon that mesenchymal stem cells could improve distal injury organ healing without cell migration and cell differentiation indicates that the cell exosomes play an important role in tissue and organ repair [ 5 – 7 ]. In addition to cell differentiation and proliferation, the secreted proteins also represent an important extension of the stem cells' function [ 8 , 9 ].…”

Proteomics Approach Highlights Early Changes in Human Fibroblasts-Pancreatic Ductal Adenocarcinoma Cells Crosstalk

“…We employed protocols without FBS supplement because it has been shown that low serum content conditions in combination with reduced biomechanical input lower the fibroblast activation state [ 24 ]. Moreover, the large number of proteins present in serum hampers in-depth proteomic analysis [ 25 ].…”

Secretomes as an emerging class of bioactive ingredients for enhanced cosmeceutical applications

“…Importantly, several authors highlighted the urgency to properly characterize secretomes composition and relate the secreted proteins to their corresponding regenerative functions. 15,18,19,71 Variations in benefits can arise due to many factors such as tissue source, 13 culture conditions and materials used in conditioned media production, 19,20,22,72 secretomes isolation methods 19 and protein detection methods 71 amongst many others. UC-MSCs reportedly produce proteins with a strong correlation to skin renewal as compared with ADSC or BM-MSCs.…”