The mesenchymal stromal cells (MSCs) have become of great interest for cell therapy because of its potential to differentiate and replenish specialized tissues. More recently, such interest has increased significantly due to the discovery that MSCs are capable of secreting a multitude of mediators to stimulate the regeneration of injured tissues in situ and immunomodulators. Therefore, MSCs may be considered both as a therapeutic product itself, such as a biofactory various relevant proteins from the standpoint of therapeutic and biotechnological. To meet these growing demands, both applications require the development of high-throughput processes under defined culture conditions, reproducible, cost-effective, allowing to obtain products with appropriate identity, strength, purity and safety. Therefore, the principal objectives of this study are to establish an efficient expansion process both in terms of growth and recovery of mesenchymal stromal cells and characterize the secretome, or the complement of proteins secreted by these cells during expansion, in order identifying possible relevant proteins with therapeutic applications. For the development of an efficient expansion process will evaluate many types of bioreactors (stirred tank, fixed bed, hollow-fibers). The growth and cell metabolism will be identified during the expansion process. The analysis of the maintenance of the biological properties (inhibition of proliferation and differentiation of lymphocytes) and immunophenotypic cells will be evaluated after cell recovery. To characterize quantitatively and the set of proteins secreted by MSCs during the expansion process, a quantitative approach will be used in marking cell culture with stable isotopes (SILAC), together with fractionation of intact proteins and analysis by mass spectrometry High resolution liquid chromatography coupled to (LC-MS/MS). Thus, by analyzing the various types of bioreactors is expected to introduce an efficient expansion of MSCs with high recovery functional and enabling its wide therapeutic applicability and identify secreted proteins involved in important trophic function of MSCs in cellular therapy and which may represent an interesting by-product of these cells in vitro.
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