CD133 mesenchymal cells were enriched using magnetic microbead anti-CD133 antibody from bone marrow mononuclear cells (BMMNCs). Flow cytometry and immunocytochemistry analysis using specific antibodies revealed that these cells were essentially 89 ± 4% CD133+ and 8 ± 5% CD34+. CD133+/CD34+ BMMNCs secrete important bioactive proteins such as cardiotrophin-1, angiogenic and neurogenic factors, morphogenetic proteins, and proinflammatory and remodeling factors in vitro. Single intracoronary infusions of autologous CD133+/CD34+ BMMNCs are effective and reduce infarct size in patients as analyzed by Tc99m MIBI myocardial scintigraphy. The majority of patients were treated via left coronary artery. Nine months after cell therapy, 5 out of 8 patients showed a net positive response to therapy in different regions of the heart. Uptake of Tc99 isotope and revitalization of the heart area in inferoseptal region are more pronounced (P = 0.016) as compared to apex and anterosptal regions after intracoronary injection of the stem cells. The cells chosen here have the properties essential for their potential use in cell therapy and their homing can be followed without major difficulty by the scintigraphy. The cell therapy proposed here is safe and should be practiced, as we found, in conjunction with scintigraphic observation of areas of heart which respond optimally to the infusion of autologous CD133+/CD34+ BMMNCs.

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Objective: Тo analyses the in vitro comportment of the CD133 + cells and their profile for cytokines secretion. Methods: Bone marrow samples were obtained from 5 healthy individuals. Mesenchymal stem cells CD133 + extracted by magnetic bead from human bone marrow mononuclear cells (BMMNCs). BMMNCs were isolated by density-gradient centrifugation over Ficoll-400. Isolated CD133+ cells were plated on 0.2% gelatin-coated wells in the presence of free culture medium for cytokines analysis and with rich culture medium for spherical stem cell cluster generation. The proteins detected by protein array from the three independent cell preparations were considered as bioactive proteins. Results: Isolated CD133 + cells using magnetic bead present more than 87±6% and 8±5% CD34 + cells as assessed by flow cytometry and differentiate into adherent cells. CD133 + cells generate spheroid cell clusters in rich culture medium. Biological classification of the bioactive proteins secreted by the primo culture of CD133 + / CD34 + BMMNCs after 36h in conditioned culture medium showed the presence of several categories of cytokines. Among these, the cardiac hypertrophic factor, the pro-angiogenic factors, the pro-inflammatory factors, wound healing factors such as MMPs-TIMPs, the neurophilic factors, the morphogenetic proteins and hematopoietic growth factors can be mentioned. Conclusion: Our results indicate that CD133 + extracted from BMMNCs secretes important bioactive proteins. The multiple properties of these cytokines undoubtedly offer many therapeutic advantages.