Dataset Information

Proteomic analysis of extracellular vesicles and conditioned medium from human Adipose-derived Stem/Stromal Cells and Dermal Fibroblasts

ABSTRACT: Conditioned medium (CM) and extracellular vesicles (EV) from human Adipose-derived Stem/stromal cells (ASC) and Dermal fibroblasts (DF) can represent valid cell substitutes in regenerative medicine and other therapeutic applications. Whether one of the two cell products should be preferred over the other is still under debate and no direct comparison of their proteome has been reported yet. Here, we apply a comprehensive quantitative proteomics approach to explore the protein composition of conditioned medium and EV obtained from the two cell types. We identified 1977 proteins by LC-MS/MS proteomic analysis. Unsupervised clustering analysis and PCA clearly distinguished CM and EV as separate groups, regardless the cell source. We identified 68 and 201 proteins that were more abundant or exclusively detectable in CM and EV, respectively. CM were enriched in proteins of endoplasmic reticulum, Golgi apparatus and lysosomes, whereas EV contained a large amount of GTPases, ribosome and translation-related factors. Moreover, the analysis of ASC and DF secretomes (both CM and EV) revealed the presence of cell type-specific factors. ASC-CM and -EV carried factors involved in ECM organization (hyaluronan and glycosaminoglycan metabolism) and immunological regulation (e.g. macrophage and IkB/NFkB signaling regulation), respectively. On the other hand, DF-CM and –EV were both enriched in epithelium development associated factors, whilst DF-CM in proteins involved in cellular processes regulation and -EV in Wnt signaling factors. In conclusion, this comprehensive proteomic analysis of the secretome components of ASC and DF, provides evidence of a different protein composition between CM and EV and of the presence of cell type-specific bioactive mediators suggesting their specific future use as advanced therapy medicinal products.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Primary Cell

SUBMITTER: Annapaola Andolfo

LAB HEAD: Anna Teresa Brini

PROVIDER: PXD020219 | Pride | 2020-12-23

REPOSITORIES: Pride

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Proteomic analysis of extracellular vesicles and conditioned medium from human adipose-derived stem/stromal cells and dermal fibroblasts.

Niada Stefania S Giannasi Chiara C Magagnotti Cinzia C Andolfo Annapaola A Brini Anna Teresa AT

Journal of proteomics 20201210

Conditioned medium (CM) and extracellular vesicles (EV) from Adipose-derived Stem/stromal cells (ASC) and Dermal fibroblasts (DF) represent promising tools for therapeutic applications. Which one should be preferred is still under debate and no direct comparison of their proteome has been reported yet. Here, we apply quantitative proteomics to explore the protein composition of CM and EV from the two cell types. Data are available via ProteomeXchange (identifier PXD020219). We identified 1977 pr ...[more]

PMID: 33309826

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Project description:Adipose tissue is a plentiful and easily accessible source of mesenchymal stem cells that have been shown to be multi potent and possibly have immuno suppressive capacity(Zuk, Zhu et al. 2002; McIntosh, Zvonic et al. 2006; Prichard, Reichert et al. 2008). Several studies have identified hypoxia and the molecular effector of reduced oxygen tension, HIF1Î± to be essential in the differentiation processes, stem cell proliferation and survival as well as in maintenance of stem cell characteristics (Lennon, Edmison et al. 2001; Goda, Ryan et al. 2003; Wang, Fermor et al. 2005; Lin, Lee et al. 2006; Malladi, Xu et al. 2007; Ohnishi, Yasuda et al. 2007). The fact that oxygen has a central role in cell development and metabolism makes the regulation of oxygen tension a valuable tool in the attempt of exploiting stem cells to their full potential. In the current study we investigate the transcriptional changes caused by two weeks of monolayer hypoxic expansion of human adipose tissue derived stem cells (ASCs). Application of the XVivo hypoxic workstation (BioSpherix, Redfield, NY) made it possible to conduct the expansion of six ASC lines at four different hypoxic conditions in parallel covering the range from mild (15% oxygen) to full hypoxic (1% oxygen) conditions. In this comprehensive investigation the Illumina bead microarray platform was employed to give a profound and new insight in the molecular changes effectuated by hypoxic expansion. The findings of this study demonstrate the significance of biologic variation in the transcriptional response to mild hypoxic expansion and display evidence that full hypoxic growth conditions at 1% oxygen tension selects for a more prolific cell population of reduced chondrogenic potential. Keywords: hypoxia response To investigate the transcriptional response to hypoxia, adipose tissue derived stem cells (ASCs) from six female individuals were cultured for 14 days at different oxygen tensions (ambient, 15%, 10%, 5% and 1%). Gene expression levels at each oxygen tension were compared to ambient conditions (Ambient oxygen tension) using Illumina Human-6 v2.0 microarrays.

			Action	DRS
	ASC-CM1a.mgf	Mgf
	ASC-CM1a.raw	Raw
	ASC-CM1b.mgf	Mgf
	ASC-CM1b.raw	Raw
	ASC-CM1c.mgf	Mgf