Two Dimensional Gel Electrophoresis Analysis of Mesenchymal Stem Cells

Monique Provansal (1) , Christian Jorgensen (2) , Sylvain Lehmann (1, 3) , S. Roche (1, 3)

1 IGH - Institut de génétique humaine
2 Cellules Souches, Plasticité Cellulaire, Médecine Régénératrice et Immunothérapies (IRMB)
3 PPM - Pôle Protéome de Montpellier

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.

Mots clés: Mesenchymal stem cell; 2D-PAGE; Western blotting; Silver staining; Proteomics; 2DE electrophoresis

Domaines: Génomique, Transcriptomique et Protéomique [q-bio.GN]; Biochimie [q-bio.BM]

Monique Provansal, Christian Jorgensen, Sylvain Lehmann, S. Roche. Two Dimensional Gel Electrophoresis Analysis of Mesenchymal Stem Cells. Mesenchymal Stem Cell Assays and Applications, 698, pp.431-442, 2011, Methods in Molecular Biology, ⟨10.1007/978-1-60761-999-4_31⟩. ⟨hal-00616455⟩