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OJVRTM
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.
ABSTRACT
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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