title: Two dimensional gel electrophoresis analysis of mesenchymal stem cells.
abstract: Proteomic analysis is a powerful tool to follow physiological modifications and phenotypes of mesenchymal stem cells (MSC). This approach generates informative data on expression and post-translational modifications of proteins which are of interest to assess the true potential of MSC in regenerative medicine. No matter the technologies used, proteomic analysis is always a challenge as the proteome is extremely diverse (in terms of constituents and concentrations), is changing with time, and is highly sensitive to pre-analytical conditions. In the framework of a European project (GENOSTEM http://www.genostem.org/), we have set up a multisite two dimensional gel electrophoresis (2DE) proteomic comparison of MSC. The goal is to compare cells from different origins, to follow their differentiation and to ultimately define a specific MSC proteomic signature. One important initial task is the optimization of 2DE protocols such that they are robust enough to be used in a multisite project. In this chapter, we detail these protocols which can be used not only for MSC but also for other cells in culture.
authors: CNRS, Institut de Génétique Humaine UPR1142, Montpellier, France.
authors: Monique
authors: M
authors: Provansal
authors: Christian
authors: C
authors: Jorgensen
authors: Sylvain
authors: S
authors: Lehmann
authors: Stéphane
authors: S
authors: Roche
mesh_terms: D015153:Blotting, Western; D002457:Cell Extracts; D004247:DNA / Q000302:isolation & purification; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007525:Isoelectric Focusing; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D008970:Molecular Weight; D011506:Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D016622:Silver Staining
chemical_list: D002457:Cell Extracts; D011506:Proteins; D004247:DNA