Overlapping of mononuclear cells derived from bone marrow in rats' intervertebral discs: an in vitro study

In this study, bone marrow mononuclear cells (BM-MNC) derived from rats were used in order to promote intervertebral disc regeneration. These cells were isolated after centrifugation in a Ficoll-Paque™ PLUS density gradient and then placed in plastic dishes to proliferate during a period of 14 days. The BM-MNCs were previously labeled with the fluorescent membrane marker Chloromethyl-benzamidodialkylcarbocyanine (CM-DIL), and thereafter were implanted in rats' intervertebral discs explants as an in vitro experimental model. Daily analyses of the cells under a fluorescence microscope revealed morphological changes, which assumed a thin and elongated shape similar to cells that originally form the annulus fibroses. Histopathological analysis demonstrated the presence of mononuclear cells interspersed within collagen fibers. The presence of viable cells, in which were found morphological changes and their disposal in the same pattern of the layers that originate the annulus fibrosus, is an indicator that they engrafted and proliferated on the intervertebral disc. Therefore, morphological changes presented by these cells indicate that they presented mesenchymal stem-like cell characteristics.

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Bibliographic Details
Main Authors: Fontes,Erica Batista, Borges,Andréa Pacheco Batista, Pippi,Ney Luis, Rosa,Maurício, Sprada,Arícia, Togni,Monique
Format: Digital revista
Language:English
Published: Universidade Federal de Santa Maria 2010
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782010000400024
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Description
Summary:In this study, bone marrow mononuclear cells (BM-MNC) derived from rats were used in order to promote intervertebral disc regeneration. These cells were isolated after centrifugation in a Ficoll-Paque™ PLUS density gradient and then placed in plastic dishes to proliferate during a period of 14 days. The BM-MNCs were previously labeled with the fluorescent membrane marker Chloromethyl-benzamidodialkylcarbocyanine (CM-DIL), and thereafter were implanted in rats' intervertebral discs explants as an in vitro experimental model. Daily analyses of the cells under a fluorescence microscope revealed morphological changes, which assumed a thin and elongated shape similar to cells that originally form the annulus fibroses. Histopathological analysis demonstrated the presence of mononuclear cells interspersed within collagen fibers. The presence of viable cells, in which were found morphological changes and their disposal in the same pattern of the layers that originate the annulus fibrosus, is an indicator that they engrafted and proliferated on the intervertebral disc. Therefore, morphological changes presented by these cells indicate that they presented mesenchymal stem-like cell characteristics.