Neurosphere generation from dental pulp of adult rat incisor
Neurosphere generation from dental pulp of adult rat incisor
复制标题
DOI：
10.1111/j.1460-9568.2008.06026.x
复制
发表时间：
2008-02-01
影响因子：
3.4
通讯作者：
Ogiuchi, Hideki
中科院分区：
文献类型：
--
作者：
Sasaki, Ryo;Aoki, Shunsuke;Ogiuchi, Hideki
牙髓是一种潜在的细胞来源，可用于各种神经系统疾病的细胞替代疗法。在这里，我们报告，成年大鼠牙髓细胞有能力形成神经球时，培养在无血清培养基上的超亲水板。小球体内的细胞继续生长，牙髓来源的细胞生成大球体。球体的形成依赖于外源性碱性成纤维细胞生长因子，但不依赖于表皮生长因子，牙髓来源的球体的形成和生长受到转化生长因子β的负调控。将从球体解离的细胞接种在粘附性基底上导致分化为Tuj1和MAP 2阳性神经元细胞。牙髓衍生球的三维结构的分析表明，它们包含巢蛋白阳性祖细胞，Tuj1阳性神经元细胞和S100阳性胶质细胞。我们发现，球体包含CD81（TAPA 1）和巢蛋白双阳性细胞，并确定了一个小的人口CD81和巢蛋白双阳性细胞在牙髓的成牙本质细胞层。流式细胞仪分析显示，CD81阳性细胞富集在球相比，牙髓组织。溴脱氧尿苷（BrdU）染色显示，巢蛋白和BrdU阳性细胞仅位于牙髓的根尖部，根尖部产生大量的大尺寸的球体。这些数据表明，CD81和巢蛋白双阳性细胞定位于牙髓的根尖部的成牙本质细胞层可能具有生长和形成神经球的能力。
Dental pulp is a potential source of cells that can be used in cell replacement therapy for various nervous system disorders. Here we report that adult rat dental pulp cells have the ability to form neurospheres when cultured in serum-free culture medium on super-hydrophilic plates. The cells within small spheres continued to grow, and the dental pulp-derived cells generated large spheres. Sphere formation was dependent on exogenously supplied basic-fibroblast growth factor, but not on epidermal growth factor, and the formation and growth of dental pulp-derived spheres were negatively regulated by transforming growth factor-beta. Plating cells that were dissociated from spheres on an adhesive substrate resulted in differentiation into Tuj1- and MAP2-positive neuronal cells. Analysis of the three-dimensional structure of dental pulp-derived spheres shows that they contained nestin-positive progenitors, Tuj1-positive neuronal cells and S100-positive glial cells. We found that spheres contained CD81 (TAPA1) and nestin double-positive cells, and identified a small population of CD81 and nestin double-positive cells in the odontoblast layer of the dental pulp. Flow cytometric analysis showed that CD81-positive cells were enriched in the spheres compared with the dental pulp tissue. Bromodeoxyuridine (BrdU) staining showed that nestin- and BrdU-positive cells were located only in the apical portion of the dental pulp, and the apical portion produced a large number of large-sized spheres. These data suggest that the CD81 and nestin double-positive cells localized in the odontoblast layer of the apical portion of the dental pulp may have the ability to grow and form neurospheres.