title: Immunophenotypic, immunocytochemistry, ultrastructural, and cytogenetic characterization of mesenchymal stem cells from equine bone marrow.
abstract: The aim of this study was to isolate, culture, and characterize mesenchymal stem cells (MSCs) from horse bone marrow (BM) using the techniques of flow cytometry, immunocytochemistry, cytogenetics, and electron microscopy. Immunophenotypic analysis revealed the presence of MSCs with high expression of the CD90 marker, lower expression of the CD44 marker, and absent expression of the CD34 marker. In assays of differentiation, the positive response to osteogenic (OST), chondrogenic (CDG), and adipogenic (ADP) differentiation signals was observed and characterized by deposition of calcium-rich extracellular matrix (OST), proteoglycans and collagen II (CDG) and intracellular deposition of fat drops (ADP). In immunocytochemical characterization, MSCs were immunopositive for CD44, vimentin, and PCNA, and they were negative for CD13. In the ultrastructural analysis of MSCs, the most outstanding characteristic was the presence of rough endoplasmic reticulum with very dilated cisterns filled with a low electrodensity material. Additionally, MSCs had normal karyotypes (2n = 64) as evidenced by cytogenetic analysis, and aneuploidy in metaphase was not observed. The protocols for isolating, culturing, and characterizing equine MSCs used in this study were shown to be appropriate for the production of a cell population with a good potential for differentiation and without aneuploidy that can be used to study future cellular therapies.
authors.affiliation: Department of Veterinary Clinics, College of Veterinary Medicine and Animal Science, São Paulo State University, Botucatu, São Paulo, Brazil.
authors.forename: Leandro
authors.initials: L
authors.lastname: Maia
authors.forename: Fernanda C
authors.initials: FC
authors.lastname: Landim-Alvarenga
authors.forename: Ligia S L Silveira
authors.initials: LS
authors.lastname: Da Mota
authors.forename: Marjorie
authors.initials: M
authors.lastname: De Assis Golim
authors.forename: Reneé
authors.initials: R
authors.lastname: Laufer-Amorim
authors.forename: Bruna
authors.initials: B
authors.lastname: De Vita
authors.forename: Danielle Jaqueta
authors.initials: DJ
authors.lastname: Barberini
authors.forename: Amanda Jeronimo
authors.initials: AJ
authors.lastname: Listoni
authors.forename: Carolina Nogueira
authors.initials: CN
authors.lastname: De Moraes
authors.forename: Marta Cristina Thomas
authors.initials: MC
authors.lastname: Heckler
authors.forename: Rogério Martins
authors.initials: RM
authors.lastname: Amorim
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000032:analysis*; D001853:Bone Marrow; D002478:Cells, Cultured; D003582:Cytogenetics; D005434:Flow Cytometry; D006736:Horses; D007150:Immunohistochemistry; D016130:Immunophenotyping; D007621:Karyotyping; D059630:Mesenchymal Stem Cells / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure*; D008854:Microscopy, Electron
chemical_list: D015703:Antigens, CD