Collection and isolation of mesenchymal stem cells from equine bone marrow.

摘要

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 waswashed 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.