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Immunophenotypic, immunocytochemistry, ultrastructural, and cytogenetic characterization of mesenchymal stem cells from equine bone marrow

2013; Wiley; Volume: 76; Issue: 6 Linguagem: Inglês

10.1002/jemt.22208

1097-0029

Leandro Maia, Fernanda da Cruz Landim‐Alvarenga, Lígia Souza Lima Silveira da Mota, Márjorie de Assis Golim, Renée Laufer Amorim, Bruna De Vita, Danielle Jaqueta Barberini, Amanda J. Listoni, Carolina Nogueira de Moraes, Marta Cristina Thomas Heckler, Rogério Martins Amorim,

Electrospun Nanofibers in Biomedical Applications

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 (2 n = 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. Microsc. Res. Tech. 76:618–624, 2013 . © 2013 Wiley Periodicals, Inc.