Sphere therapy for corneal endothelium deficiency in a rabbit model
Sphere therapy for corneal endothelium deficiency in a rabbit model
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DOI：
10.1167/iovs.05-0251
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发表时间：
2005-09-01
影响因子：
4.4
通讯作者：
Amano, S
中科院分区：
文献类型：
--
作者：
Mimura, T;Yamagami, S;Amano, S
目的.从兔角膜内皮细胞（CE）中分离前体细胞，并将其用于治疗CE缺陷的兔模型。进行成球测定以从兔CE分离前体细胞。用免疫细胞化学方法检测神经细胞和间充质细胞在球形集落及其后代中的标志物表达。通过测量电位差和短路电流来评价CE片材的泵功能。将通过球体形成测定从兔CE获得的前体注射到眼前房中，之后眼睛向下（即，CE向上）的位置保持24小时，以允许通过重力进行附着（球形眼睛向下组）。眼球向下组和对照组，术后观察28天，进行组织学和荧光显微镜检查。兔CE形成原代和次级球形集落。后代表达α-平滑肌肌动蛋白，巢蛋白，和神经标记，并表现出CE样六边形的形状和足够的运输活动。术后第3、7、14、21、28天，球形眼向下组的平均角膜厚度显著小于其他对照组（P < 0.05）。角膜后弹力层上可见CE样六角形细胞，角膜水肿明显减轻。DiI标记的细胞仅在球形眼向下组中散布于角膜后表面。通过成球测定分离来自兔CE的前体。兔CE衍生球治疗是兔CE缺乏模型的有效治疗。
PURPOSE. To isolate precursor cells derived from rabbit corneal endothelium (CE) and to use them for the treatment of CE deficiency in a rabbit model.METHODS. A sphere-forming assay was performed to isolate precursor cells from rabbit CE. Immunocytochemistry was used to examine marker expressions of neural and mesenchymal cells in the sphere colonies and their progenies. The pump function of the CE sheet was evaluated by measurement of the potential difference and short circuit current. Precursors obtained from rabbit CE by a sphere-forming assay were injected into the anterior chamber of the eye, after which an eye-down (i.e., CE up) position was maintained for 24 hours to allow attachment by gravitation ( sphere eye-down group). The sphere eye-down and control groups, observed for 28 days after surgery, underwent histologic and fluorescence microscopic examinations.RESULTS. Rabbit CE formed primary and secondary sphere colonies. The progeny expressed alpha-smooth muscle actin, nestin, and neural markers and showed a CE-like hexagonal shape and adequate transport activity. Mean corneal thickness in the sphere eye-down group was significantly less than in the other control groups 3, 7, 14, 21, and 28 days ( P < 0.05) after surgery. CE-like hexagonal cells were detected on Descemet's membrane, and corneal edema was substantially suppressed. DiI-labeled cells were spread over the rear corneal surface in the sphere eye-down group only.CONCLUSIONS. Precursors from rabbit CE were isolated by a sphere-forming assay. Rabbit CE-derived sphere therapy is an effective treatment in a rabbit CE deficiency model.