Culture and characterisation of equine peripheral blood mesenchymal stromal cells |
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| Authors: | Jan H Spaas Catharina De Schauwer Pieter Cornillie Evelyne Meyer Ann Van Soom Gerlinde R Van de Walle |
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| Institution: | 1. Department of Comparative Physiology and Biometrics, Faculty of Veterinary Medicine, Ghent University, B-9820 Merelbeke, Belgium;2. Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, B-9820 Merelbeke, Belgium;3. Department of Morphology, Faculty of Veterinary Medicine, Ghent University, B-9820 Merelbeke, Belgium;4. Department of Pharmacology, Toxicology and Biochemistry, Faculty of Veterinary Medicine, Ghent University, B-9820 Merelbeke, Belgium;1. Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, NC 27607, United States;2. Department of Molecular Biomedical Sciences, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, NC 27607, United States;3. Center for Comparative Medicine and Translational Research, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, NC 27607, United States;1. Virological Diagnostic Area, Experimental Zooprofilactic Institute of Sicily “A. Mirri”, Palermo, Italy;2. Department of Veterinary Science, Polo Universitario Annunziata, University of Messina, Messina, Italy;1. Department of Anesthesiology, Daejeon St. Mary''s Hospital, The Catholic University of Korea, Daejeon, Republic of Korea;2. Department of Surgery, Daejeon St. Mary''s Hospital, The Catholic University of Korea, Daejeon, Republic of Korea;1. Department of Clinical Veterinary Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan;2. Department of Basic Veterinary Medicine, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan;3. Department of Biomaterials, Institute for Frontier Medical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8507, Japan;1. Universidad de Concepción, Campus Chillan, Faculty of Veterinary Science, Department of Animal Science, Laboratorio de Biotecnología Animal, Chile;2. Universidad de Concepción, Campus Chillan, Faculty of Veterinary Sciences, Department of Animal Pathology, Chile |
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| Abstract: | Although the use of mesenchymal stromal cells (MSCs) for the treatment of orthopaedic injuries in horses has been reported, no official guidelines exist that classify a particular cell as an equine MSC. Given the limited characterisation of peripheral blood (PB)-derived equine MSCs in particular, this study aimed to provide more detailed information in relation to this cell type. Mesenchymal stromal cells were isolated from equine PB samples and colony forming unit (CFU) assays as well as population doubling times (PDTs) (from P0 to P10) were performed.Two types of colonies, ‘fingerprint’ and dispersed, could be observed based on macroscopic and microscopic features. Moreover, after an initial lag phase (as indicated by a negative PDT at P0 to P1) the MSCs divided rapidly as indicated by a positive PDT at all further passages. Immunophenotyping was carried out with trypsin- as well as with accutase-detached MSC to evaluate potential trypsin-sensitive epitope destruction on particular antigens. Isolated MSC were positive for CD29, CD44, CD90 and CD105, and negative for CD45, CD79α, MHC II and a monocyte/macrophage marker, irrespective of the cell detaching agent used. Trilineage differentiation of the MSCs towards osteoblasts, chondroblasts and adipocytes was confirmed using a range of histochemical stains. |
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