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Collection, processing and freezing of equine bone marrow cells

Bastos, Fernanda Z., Barussi, Fernanda C.M., Santi, Thasla F., Vieira, Bianca P., Senegaglia, Alexandra C., Cruz, Fernanda F., Michelotto, Pedro V.
Cryobiology 2017
bone marrow, bone marrow cells, cell viability, cryopreservation, elderly, freezing, frozen storage, heparin, horses, nitrogen, sternum, syringes
There is no consensus on aspects of equine bone marrow collection and processing. The study aimed to describe the collection of large volumes of bone marrow from horses of advanced age, with emphasis on bone marrow mononuclear cells (BMMCs) recovery and viability after cryopreservation. Fourteen horses, aged 3–24 years, were divided into three experiments. E1 studied the feasibility of collecting 200 mL from the sternums of horses of advanced age; E2 examined the number of cells obtained from the first and last syringe of each puncture; and E3 investigated the influence of heparin concentration on the prevention of cell aggregation, and cell viability after freezing in liquid nitrogen. Bone marrow aspirations were done with syringes pre-filled with Iscove's modified Dulbecco's medium and different concentrations of sodium heparin. BMMCs were counted, cell viability was determined, and samples were frozen. Bone marrow collection from the sternum is safe, even at large volumes and from horses of advanced age, and the number of cells recovered decreases with successive aspirations (p < 0.0001). Heparin concentration influenced cell aggregation, and recovered cells continued to be commercially viable after 150 days in frozen storage.