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首页> 外文期刊>Cell and Tissue Research >Isolation and characterization of cluster of differentiation 9-positive ependymal cells as potential adult neural stem/progenitor cells in the third ventricle of adult rats
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Isolation and characterization of cluster of differentiation 9-positive ependymal cells as potential adult neural stem/progenitor cells in the third ventricle of adult rats

机译:分化9阳性突出细胞簇的分离与表征作为成年大鼠第三脑室中的潜在成年神经茎/祖细胞

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Ependymal cells located above the ventricular zone of the lateral, third, and fourth ventricles and the spinal cord are thought to form part of the adult neurogenic niche. Many studies have focused on ependymal cells as potential adult neural stem/progenitor cells. To investigate the functions of ependymal cells, a simple method to isolate subtypes is needed. Accordingly, in this study, we evaluated the expression of cluster of differentiation (CD) 9 in ependymal cells by in situ hybridization and immunohistochemistry. Our results showed that CD9-positive ependymal cells were also immunopositive for SRY-box 2, a stem/progenitor cell marker. We then isolated CD9-positive ependymal cells from the third ventricle using the pluriBead-cascade cell isolation system based on antibody-mediated binding of cells to beads of different sizes and their isolation with sieves of different mesh sizes. As a result, we succeeded in isolating CD9-positive populations with 86% purity of ependymal cells from the third ventricle. We next assayed whether isolated CD9-positive ependymal cells had neurospherogenic potential. Neurospheres were generated from CD9-positive ependymal cells of adult rats and were immunopositve for neuron, astrocyte, and oligodendrocyte markers after cultivation. Thus, based on these findings, we suggest that the isolated CD9-positive ependymal cells from the third ventricle included tanycytes, which are special ependymal cells in the ventricular zone of the third ventricle that form part of the adult neurogenic and gliogenic niche. These current findings improve our understanding of tanycytes in the adult third ventricle in vitro.
机译:据认为,位于横向,第三和第四脑室和第四脑室和脊髓的心室区域上方的突出细胞,形成成人神经源性的部分。许多研究专注于突出突细胞作为潜在的成年神经茎/祖细胞。为了探讨突变细胞的功能,需要一种分离亚型的简单方法。因此,在本研究中,我们通过原位杂交和免疫组化评估了在突突细胞中的分化(CD)9簇的表达。我们的结果表明,CD9阳性突出细胞也是Sry-Box 2,茎/祖细胞标记的免疫阳性。然后,我们使用基于抗体介导的细胞与不同尺寸的珠子的抗体介导的细胞分离系统与不同尺寸的珠子的珠子分离CD9阳性突出细胞,以及与不同网眼尺寸的筛的分离。结果,我们成功地将CD9阳性群体分离出来自第三脑室的86%纯度的突变细胞。我们接下来测定分离的CD9阳性突突细胞是否具有神经疏松潜力。从成年大鼠的CD9阳性突变细胞产生神经球,并为培养后的神经元,星形胶质细胞和少突胶质细胞标志物的免疫键。因此,基于这些发现,我们表明来自第三心室的分离的CD9阳性突突细胞包括Tanycytes,其是第三脑室的心室区域中的特殊突变细胞,其形成成人神经源性和胶质原性的部分。这些目前的调查结果改善了我们对体外成人第三心室的坦宁细胞的理解。

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