Aislamiento, viabilidad y rendimiento de células mononucleares de médula ósea de conejos

Abstract

The aim of this study was to establish a cellular isolation protocol and evaluate the yield and viability of bone marrow mononuclear cells in rabbits. Thirty-two male adult New Zealand rabbits were used with 3.06 ± 0.24 Kg mean weight. A volume of 2.0 mL bone marrow was aspirated from the humerus of each animal. Samples were diluted in Dulbecco phosphate buffered saline (DPBS) and submitted to density gradient using Ficoll-Ipaque (1.077 g/mL) at a 2:1 rate. Subsequently, the material was centrifuged at 495g during 30 minutes, at 15 oC. The yield and cell viability were determined in Neubauer haemocytometer chamber, using trypan blue vital staining. A mean yield [±SD] of 6.65 [±4.18] x106 cells/mL was obtained and the mean viability [±SD] was 92.66 [±4.35] %. The protocol using density gradient was effective for the isolation, yield and viability of bone marrow mononuclear cells in rabbits in order to use them in cell therapy.