PXD002217

PXD002217 is an original dataset announced via ProteomeXchange.

Title	Variations of secretome profiles according to conditioned medium preparation: the example of human mesenchymal stem cell derived adipocytes
Description	One challenging point in analysing 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 one dimensional 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.
HostingRepository	PRIDE
AnnounceDate	2015-07-03
AnnouncementXML	Submission_2015-07-03_13:16:25.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Thibaut Léger
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	deamidated residue; monohydroxylated residue; acetylated residue; iodoacetamide derivatized residue; phosphorylated residue
Instrument	LTQ Orbitrap Velos

Revision	Datetime	Status	ChangeLog Entry
0	2015-05-22 08:06:47	ID requested
⏵ 1	2015-07-03 13:16:26	announced

Clabaut A, Grare C, L, é, ger T, Hardouin P, Broux O, Variations of secretome profiles according to conditioned medium preparation: The example of human mesenchymal stem cell-derived adipocytes. Electrophoresis, 36(20):2587-93(2015) [pubmed]

curator keyword: Biomedical

submitter keyword: Adipocyte/Conditioned medium/Mesenchymal stem cell /Secretome

Odile Broux
contact affiliation	PMOI, ULCO, Bassin Napoléon, BP 120, F-62327 Boulogne-sur-Mer, France Phone: +33(0)321994521. Fax: +33(0)321994524.
contact email	broux@univ-littoral.fr
lab head
Thibaut Léger
contact affiliation	Institut Jacques Monod
contact email	leger@ijm.univ-paris-diderot.fr
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD002217
     1. Label: PRIDE project
     2. Name: Variations of secretome profiles according to conditioned medium preparation: the example of human mesenchymal stem cell derived adipocytes