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Online Journal of Veterinary Research©
Volume 19(3): 216-223, 2015.
Collection and isolation of mesenchymal stem cells from equine bone marrow.
Luiz Fernando de Oliveira Varanda1, Julia de Miranda Moraes1 , Juliana Vieira Flores Sales1, Ana Lourdes Arrais de Alencar Mota1, André Rodrigues da Cunha Barreto-Vianna1, Roberta Ferro de Godoy2, Eduardo Maurício Mendes de Lima1*
1Faculdade de Agronomia e Medicina Veterinária, Universidade de Brasília (UnB), ICC Sul, Campus Universitário Darcy Ribeiro, Brasília, Brazil. 2Institute of Orthopaedics and Musculoskeletal Science, National Orthopaedics Hospital, University College of London, Brockley Hill Stanmore, HA7 4LP.United Kingdom.
Oliveira Varanda LF, Miranda Moraes J, Flores Sales JV, Arrais de Alencar Mota AL, Cunha Barreto-Vianna AR, Godoy RF, Mendes de Lima EM., Collection and isolation of mesenchymal stem cells from equine bone marrow, Onl J Vet Res., 19(3): 212-223, 2015. Mesenchymal stem cells have been studied for tissue repair in several species. It is a promising source for the treatment of equine musculoskeletal diseases. We describe isolation of the fraction of mononuclear cells (FMC) collected from bone marrow (BM) in standing horses. Four female and two male horses weighing ~248 kg, aged 1.5 to 10 years were used. BM was aspirated from the sternum bone between the 4th and 5th sternebra. To avoid clot formation in the medullary cavity and needle, the needle was washed with heparin solution and 10 mL of this solution was injected into the medullary cavity before aspiration. FMC was isolated from the aspirate via centrifugation and Ficoll-Hypaque concentration gradient separation. Samples were stained with trypan blue for nucleated cell count and determination of cell viability. Means of nucleated and mesenchymal stem cells obtained after isolation were 7.1x10⁷ and 710 cells/mL, respectively; mean cell viability was 86.2%. We find the procedure to be of low-cost and able to be performed routinely.
Keywords: bone marrow collection, fraction of mononuclear cells, cell therapy, equine tissue repair.
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