Yoshida Satoru, Shimmura Shigeto, Nagoshi Narihito, Fukuda Keiichi, Matsuzaki Yumi, Okano Hideyuki, Tsubota Kazuo
Cornea Center, Tokyo Dental College, Ichikawa, Chiba, Japan.
Stem Cells. 2006 Dec;24(12):2714-22. doi: 10.1634/stemcells.2006-0156. Epub 2006 Aug 3.
We report the presence of neural crest-derived corneal precursors (COPs) that initiate spheres by clonal expansion from a single cell. COPs expressed the stem cell markers nestin, Notch1, Musashi-1, and ABCG2 and showed the side population cell phenotype. COPs were multipotent with the ability to differentiate into adipocytes, chondrocytes, as well as neural cells, as shown by the expression of beta-III-tubulin, glial fibrillary acidic protein, and neurofilament-M. COP spheres prepared from E/nestin-enhanced green fluorescent protein (EGFP) mice showed induction of EGFP expression that was not originally observed in the cornea, indicating activation of the neural-specific nestin second intronic enhancer in culture. COPs were Sca-1(+), CD34(+), CD45(-), and c-kit(-). Numerous GFP(+) cells were observed in the corneas of mice transplanted with whole bone marrow of transgenic mice ubiquitously expressing GFP; however, no GFP(+) COP spheres were initiated from these mice. On the other hand, COP spheres from transgenic mice encoding P0-Cre/Floxed-EGFP as well as Wnt1-Cre/Floxed-EGFP were GFP(+), indicating the neural crest origin of COPs, which was confirmed by the expression of the embryonic neural crest markers Twist, Snail, Slug, and Sox9. Taken together, these data indicate the existence of neural crest-derived, multipotent stem cells in the adult cornea.
我们报告了神经嵴来源的角膜前体细胞（COPs）的存在，这些细胞通过单个细胞的克隆扩增形成球体。COPs表达干细胞标志物巢蛋白、Notch1、Musashi-1和ABCG2，并表现出侧群细胞表型。如β-III-微管蛋白、胶质纤维酸性蛋白和神经丝-M的表达所示，COPs具有多能性，能够分化为脂肪细胞、软骨细胞以及神经细胞。从E/nestin-增强型绿色荧光蛋白（EGFP）小鼠制备的COP球体显示出EGFP表达的诱导，而这在角膜中原本未观察到，表明培养中神经特异性巢蛋白第二内含子增强子被激活。COPs为Sca-1(+)、CD34(+)、CD45(-)和c-kit(-)。在移植了普遍表达GFP的转基因小鼠全骨髓的小鼠角膜中观察到大量GFP(+)细胞；然而，这些小鼠并未起始GFP(+) COP球体。另一方面，来自编码P0-Cre/Floxed-EGFP以及Wnt1-Cre/Floxed-EGFP的转基因小鼠的COP球体为GFP(+)，表明COPs起源于神经嵴，这通过胚胎神经嵴标志物Twist、Snail、Slug和Sox9的表达得到证实。综上所述，这些数据表明成年角膜中存在神经嵴来源的多能干细胞。
