Yamagami Satoru, Yokoo Seiichi, Mimura Tatsuya, Takato Tsuyoshi, Araie Makoto, Amano Shiro
Department of Corneal Tissue Regeneration, Tokyo University Graduate School of Medicine, Tokyo, Japan.
Ophthalmology. 2007 Mar;114(3):433-9. doi: 10.1016/j.ophtha.2006.07.042.
We identified original tissue-committed precursors with limited self-renewal capacity from human corneal stromal (HCS) cells and human corneal endothelial (HCE) cells, then tried to determine the distribution and proliferative capacity of the precursors.
Experimental study.
Eighteen human corneas from donors 56 to 68 years old.
Human corneal stromal cells were divided into groups based on distance from the center of the cornea: <6 mm (central), 6 to 8 mm (paracentral), and 8 to 10 mm (peripheral). Human corneal endothelial cells were separated into 2 groups: <7.5 mm (central) and 7.5 to 10 mm (peripheral) from the center. Each group was subjected to the sphere-forming assay using serum-free medium containing growth factors in floating culture. Sphere numbers and the proliferative capacity of spheres in adherent culture were compared among the groups.
Density and proliferative capacity of precursors from each area of HCS and HCE cells.
Primary spheres were isolated from all groups of HCS and HCE cells. The rate of primary sphere formation from peripheral HCS cells was higher than those of the other 2 groups, being 1.5-fold greater than in the paracentral cornea and 4-fold greater than in the central cornea. The rate of primary sphere formation by peripheral HCE cells was significantly higher than that by central HCE cells, being 4-fold greater than in the central cornea. There were no differences in the proliferative capacity of HCS and HCE cell spheres from the different areas after adherent culture.
All HCS and HCE cells contain a significant number of precursors, but the peripheral cells have a density of precursors higher than that of the central cells. Precursors from each area do not show differences of proliferative capacity. Our findings may in part explain changes after excimer laser treatment and may have implications for corneal transplantation procedures.
我们从人角膜基质（HCS）细胞和人角膜内皮（HCE）细胞中鉴定出具有有限自我更新能力的原始组织定向前体细胞，然后试图确定这些前体细胞的分布和增殖能力。
实验研究。
来自56至68岁供体的18个人类角膜。
人角膜基质细胞根据距角膜中心的距离分为几组：<6毫米（中央）、6至8毫米（旁中央）和8至10毫米（周边）。人角膜内皮细胞分为两组：距中心<7.5毫米（中央）和7.5至10毫米（周边）。每组在含有生长因子的无血清培养基中进行悬浮培养的成球试验。比较各组在贴壁培养中的球数和球的增殖能力。
HCS和HCE细胞各区域前体细胞的密度和增殖能力。
从所有HCS和HCE细胞组中分离出初级球。周边HCS细胞的初级球形成率高于其他两组，比旁中央角膜高1.5倍，比中央角膜高4倍。周边HCE细胞的初级球形成率显著高于中央HCE细胞，比中央角膜高4倍。贴壁培养后，不同区域的HCS和HCE细胞球的增殖能力没有差异。
所有HCS和HCE细胞都含有大量前体细胞，但周边细胞的前体细胞密度高于中央细胞。各区域的前体细胞在增殖能力上没有差异。我们的发现可能部分解释了准分子激光治疗后的变化，并可能对角膜移植手术有影响。
