Характеристика костномозгового клеточного трасплантата больных хронической сердечной недостаточностью до и после кратковременной экспозиции с эритропоэтином

Autologous bone marrow stem cells are an alternative therapy for patients with heart failure. The aim of this work was to study the phenotype and functional properties of bone marrow mononuclear cells (BM-MNCs) from patients with chronic heart failure (CHF) before and after a short-term in vitro exposure to erythropoietin. Methods. BM-MNCs were isolated using density gradient. The BM-MNC phenotype, cell cycle, and apoptosis of CD34+ cells were evaluated before and after exposure to erythropoietin with a flow cytometer. Proliferation of BM-MNCs before and after the erythropoietin exposure was evaluated in a spontaneous and stimulating test. Proliferation, migration, and angiogenic potential of EA.hy 929 cells were studied in the wound closure test and in the tubule formation test under the influence of 30% conditioned medium from BM-MNCs. Results. BM-MNCs represented a mixture of hematopoietic stem cells (HSCs), endothelial progenitor cells (EPCs) at different stages of maturation and differentiation, and mesenchymal stem cells (MSCs). Erythropoietin increased the number of CD34+ cells in the G0/G1 cell cycle phase, CD45+/EpoR+, CD31-/CD184+, CD31+/CD184+, and CD34+/CD184-, and decreased the number of CD34+/CD133+ and CD34+/EpoR. The erythropoietin exposure of BM-MNCs reduced their proliferative capacity. The BM-MNCs-conditioned medium promoted EA.hy 929 cell proliferation, migration, and formation of vascular-like structures. Conclusion. A short-term exposure of BM-MNCs delayed the resting stage of CD34+ cells, increased the pool of EPCs expressing the homing receptor while the BM-MNC conditioned medium stimulated EA.hy 929 proliferation, migration and tubule formation, which should be taken into account when selecting methods to enhance survival of cellular grafts.