Multi-User Equipment approved in grant 2013/08135-2 equipment: flow cytometer with cell Soter accessories

Abstract

With the creation of the Center for Cell-based Therapy (CTC) (FAPESP process n. 2013/08.135-2) that comprises an ambitious multidisciplinary research program whose major objective is to study molecular, cellular, and biological aspects of normal and pathological stem cells and to critically evaluate their therapeutic potential. Currently our studies are conducted in pluripotent (embryonic and iPS) stem cells and in somatic (hematopoietic, mesenchymal, and endothelial) stem cells, concentrated on two broad areas: basic research and clinical studies. In addition to these research projects, our center also develops processes of large-scale stem cell production and production of new recombinant proteins for clinical use and/or for the use in diagnostic tests used in transfusion safety and hematological diseases. This project is designed to the acquisition of the equipment FACSAria Fusion from a multipurpose and multiuser perspective to meet the demand of the Center and interested partners. The headquarter institution for the use of the equipment requested is the Regional Center of Hemotherapy of Ribeirão Preto. The applicability of this instrument is justified by the objectives proposed in the extension of the projects at the Center, once all the cells produced in our Center must be rigorously characterized. In summary, flow cytometry is based on the transportation of particles, in a hydrodynamic focusing flow, which receive the incidence of a laser. When the light goes through the cells, it is deviated by refraction and the laser may excite any chromophore for the production of fluorescence. This refracted light or fluorescence is captured by receptors and these data are processed in events via software. Therefore, cytometry offers a wide range of applicability, by the analyses of cell size and complexity, as well as surface and intracellular proteins, apart from cell cycle and proliferation among other parameters. Among the main clinical applicability we can highlight immunophenotyping which has crucial importance in the diagnosis of leukemias.The use of this cytometer will be involved in the main projects listed below:1) In the main Basic Research projects:*Establishment of human embryonic stem cells (hESC) for the use in pre-clinical studies; *Differentiation of pluripotent stem cells into blood cells; *Generation and characterization of induced pluripotent stem cells (iPS) of patients and healthy individuals; *Determination of imprinting patterns during cell reprograming; *Evaluation of the functional role of microRNAs associated with pluripotency and differentiation of pluripotent stem cells; *Evaluation of endothelial cells from different origins that move to the mesenchymal phenotype in vitro by EndMT;*Studies on the molecular mechanisms involved in the increase of immunomodulatory properties of mesenchymal cells by inflammatory stimuli. 2) In all Clinical Research projects 3) In the future projects of Research of New Therapies against cancer. (AU)