Collection,processing and freezing of equine bone marrow cells |
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| Institution: | 1. Department of Animal Science, School of Life Sciences - Pontifícia Universidade Católica Do Paraná, Curitiba, Paraná, Brazil;2. Course of Veterinary Medicine, School of Life Sciences - Pontifícia Universidade Católica Do Paraná, Curitiba, Paraná, Brazil;3. Experimental Laboratory for Cell Culture, School of Medicine- Pontifícia Universidade Católica Do Paraná, Curitiba, Paraná, Brazil;4. Laboratory of Pulmonary Investigation, Carlos Chagas Filho Biophysics Institute, Universidade Federal Do Rio de Janeiro, Rio de Janeiro, Rio de Janeiro, Brazil;1. E Singer Equine Orthopaedics and Surgery, 14 Brooklands Rd, Parkgate, Neston, Cheshire, CH646SW, UK;2. JD Wheat Veterinary Orthopedic Research Laboratory, School of Veterinary Medicine, University of California, One Shields Avenue, Davis, CA, USA;1. Centro Universitário de Mineiros – UNIFIMES, Rua 23, esquina com Av. Caiapós – Setor Aeroporto, Mineiros, GO, Brazil;2. EQUINOVA Research Group, Veterinary School, Universidade Federal de Minas Gerais (UFMG), Av. Antônio Carlos 6627, campus Pampulha, Belo Horizonte, MG 31270-901, Brazil;3. Conselho Nacional de Desenvolvimento Científico e Tecnológico, CNPq, SHIS, Edifício Santos Dumont, Lago Sul, Brasília, DF 71605-001, Brazil;1. Department of Biomedical Engineering, Melbourne School of Engineering, University of Melbourne, Australia;2. U-Vet Equine Centre, Melbourne Veterinary School, Faculty of Veterinary and Agricultural Sciences, University of Melbourne, Melbourne, Australia;1. Laboratorio de Genética Bioquímica LAGENBIO, Universidad de Zaragoza, C/Miguel Servet, 177, 50013 Zaragoza, Spain;2. Servicio de Cirugía y Medicina Equina, Hospital Veterinario, Universidad de Zaragoza, C/Miguel Servet, 177, 50013 Zaragoza, Spain;3. Servicio de Cirugía Ortopédica y Traumatología. Hospital Clínico Universitario Lozano Blesa, Zaragoza, Avda. San Juan Bosco, 15, 50009 Zaragoza Spain;4. Departament de Medicina i Cirugia Animal, Universidad Autónoma de Barcelona, Edifici H, UAB, 08193 Bellaterra, Barcelona, Spain |
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| Abstract: | 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. |
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| Keywords: | Cell therapy Cryopreservation Heparin Horse Bone marrow mononuclear cells |
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