Choroid plexus ependymal cells host neural progenitor cells in the rat
Choroid plexus ependymal cells host neural progenitor cells in the rat
复制标题
作者：
Itokazu, Y;Kitada, M;Ide, C
我们之前已经证明，在移植到脊髓后，脉络膜丛上皮（修饰室管膜）细胞（cpec）分化为星形胶质细胞。在本研究中，我们检测了出生后第1天（P1）、第7天（P7）和第8周（P8W）大鼠的cpec是否可以作为神经祖细胞产生神经元和胶质细胞。在每个产后时期用大鼠第四脑室脉络膜丛全组织培养产生细胞球。p -微管蛋白III类（Tuj-1）、胶质纤维酸蛋白(GFAP)-和o4阳性细胞在分化培养基中从细胞球分化。我们制备了一种特异性标记cpec微绒毛的单克隆抗体3E6。利用该单克隆抗体，通过细胞分选仪（FACS）从P8W大鼠脉络膜丛中分离cpec。免疫细胞化学证实，facs分离的3e6标记细胞中没有来自成纤维细胞、内皮细胞、巨噬细胞或雪旺细胞的污染。在这些3e6标记的cpec培养物中形成细胞球。细胞球扩增后产生Tuj-1-(5%)、GFAP-(45%)和o4阳性细胞（0.16%）。剩下的细胞（45%）是未标记的神经或胶质标记物。一些P8W大鼠的cpec用musashi1和表皮生长因子受体（EGF-R）的谱系相关标志物进行免疫组织化学染色。此外，脑室输注EGF或成纤维细胞生长因子-2 （FGF2）使cpec中溴脱氧尿嘧啶（BrdU）阳性细胞的数量分别从0.03%（未处理）增加到1.14%（38倍，EGF）和1.03%（35倍，FGF2）。这些发现表明大鼠cpec中存在神经祖细胞。(c) 2005 Wiley-Liss, Inc。
We previously demonstrated that choroid plexus epithelial (modified ependymal) cells (CPECs) differentiated into astrocytes after grafting into the spinal cord. In the present study, we examined whether CPECs from rats at postnatal I day (P1), 7 day (P7), and 8 weeks (P8W) can function as neural progenitor cells that give rise to neurons and glial cells. Cell spheres were produced in cultures of whole tissue of the choroid plexus from the fourth ventricle of rats at each postnatal period. P-tubulin class III (Tuj-1), glial fibrillary acid protein (GFAP)-, and O4-positive cells differentiated from cell spheres in the differentiation medium. We produced a monoclonal antibody 3E6 specifically labeling microvilli of CPECs. Using this monoclonal antibody, CPECs were isolated from the choroid plexus of P8W rats by cell sorter (FACS). Immunocytochemistry confirmed that there was no contamination from fibroblasts, endothelial cells, macrophages, or Schwann cells in the FACS-isolated 3E6-labeled cells. Cell spheres formed in the cultures of these 3E6-labeled CPECs. After expansion, these cell spheres gave rise to Tuj-1- (5%), GFAP- (45%), and O4-positive cells (0.16%). The remaining cells (45%) were unlabeled neural or glial markers. Some CPECs of the P8W rat were immunohistochemically stained with lineage-associated markers of Musashi-1 and epidermal growth factor-receptor (EGF-R). In addition, infusion of EGF or fibroblast growth factor-2 (FGF2) into the ventricle increased the number of bromodeoxyuridine (BrdU)-positive cells among CPECs from 0.03% (untreated) to 1.14% (38-fold, EGF) and 1.03% (35-fold, FGF2), respectively. These findings indicate that neural progenitor cells exist among CPECs in the rat. (c) 2005 Wiley-Liss, Inc.