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.
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References
Roche, S., Provansal, M., Tiers, L., Jorgensen, C., and Lehmann, S. (2006) Proteomics of primary mesenchymal stem cells Regen Med 1, 511–7.
Tian, Q., Stepaniants, S. B., Mao, M., Weng, L., Feetham, M. C., Doyle, M. J., Yi, E. C., Dai, H., Thorsson, V., Eng, J., Goodlett, D., Berger, J. P., Gunter, B., Linseley, P. S., Stoughton, R. B., Aebersold, R., Collins, S. J., Hanlon, W. A., and Hood, L. E. (2004) Integrated genomic and proteomic analyses of gene expression in Mammalian cells Mol Cell Proteomics 3, 960–9.
Roche, S., Delorme, B., Oostendorp, R. A., Barbet, R., Caton, D., Noel, D., Boumediene, K., Papadaki, H. A., Cousin, B., Crozet, C., Milhavet, O., Casteilla, L., Hatzfeld, J., Jorgensen, C., Charbord, P., and Lehmann, S. (2009) Comparative proteomic analysis of human mesenchymal and embryonic stem cells: towards the definition of a mesenchymal stem cell proteomic signature Proteomics 9, 223–32.
Noel, D., Caton, D., Roche, S., Bony, C., Lehmann, S., Casteilla, L., Jorgensen, C., and Cousin, B. (2008) Cell specific differences between human adipose-derived and mesenchymal-stromal cells despite similar differentiation potentials Exp Cell Res 314, 1575–84.
Kastrinaki, M. C., Sidiropoulos, P., Roche, S., Ringe, J., Lehmann, S., Kritikos, H., Vlahava, V. M., Delorme, B., Eliopoulos, G. D., Jorgensen, C., Charbord, P., Haupl, T., Boumpas, D. T., and Papadaki, H. A. (2008) Functional, molecular and proteomic characterisation of bone marrow mesenchymal stem cells in rheumatoid arthritis Ann Rheum Dis 67, 741–9.
Rabilloud, T. (1998) Use of thiourea to increase the solubility of membrane proteins in two-dimensional electrophoresis Electrophoresis 19, 758–60.
Rabilloud, T. (2000) Proteome Research: Two-Dimensional Gel Electrophoresis and Identification Methods, Springer, Heidelberg.
Seo, J., Bakay, M., Chen, Y. W., Hilmer, S., Shneiderman, B., and Hoffman, E. P. (2004) Interactively optimizing signal-to-noise ratios in expression profiling: project-specific algorithm selection and detection p-value weighting in Affymetrix microarrays Bioinformatics 20, 2534–44.
Shevchenko, A., Wilm, M., Vorm, O., and Mann, M. (1996) Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels Anal Chem 68, 850–8.
Acknowledgments
This work was supported by the European Commission through the FP6 project GENOSTEM (LHSB-CT-2003-503161).
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Provansal, M., Jorgensen, C., Lehmann, S., Roche, S. (2011). Two Dimensional Gel Electrophoresis Analysis of Mesenchymal Stem Cells. In: Vemuri, M., Chase, L., Rao, M. (eds) Mesenchymal Stem Cell Assays and Applications. Methods in Molecular Biology, vol 698. Humana Press. https://doi.org/10.1007/978-1-60761-999-4_31
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DOI: https://doi.org/10.1007/978-1-60761-999-4_31
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