Koizumi Noriko, Sakamoto Yuji, Okumura Naoki, Okahara Norio, Tsuchiya Hideaki, Torii Ryuzo, Cooper Leanne J, Ban Yuriko, Tanioka Hidetoshi, Kinoshita Shigeru
Research Center for Regenerative Medicine, Doshisha University, Kyoto, Japan.
Invest Ophthalmol Vis Sci. 2007 Oct;48(10):4519-26. doi: 10.1167/iovs.07-0567.
To examine the feasibility of cultivated corneal endothelial cell transplantation in a primate model.
Monkey corneal endothelial cells (MCECs) obtained from three cynomolgus monkeys were cultivated, with subcultures grown on collagen type I carriers for 4 weeks. The corneal endothelium of the right eye of six monkeys was mechanically scraped, after which a cultivated MCEC sheet was brought into the anterior chamber of four eyes and fixed to Descemet's membrane by air. As the control, a collagen sheet without MCECs was transplanted into one eye of one monkey, and a suspension of cultivated MCECs was injected into the anterior chamber in one eye.
Cultivated MCECs produced a confluent monolayer of closely attached hexagonal cells that showed both ZO-1 and Na(+)-K(+) ATPase expression. In the early postoperative period MCEC sheets were attached to Descemet's membrane and corneal clarity was recovered. The recovered clarity was accompanied by a decrease in corneal thickness. Fluorescein DiI labeled donor corneal endothelial cells were detected on the host cornea on postoperative day 7. Six months after transplantation MCEC-transplanted corneas remained clear, and hexagonal cells were observed by in vivo specular microscopy with a density of 1992 to 2475 cells/mm(2). Control eyes showed irreversible bullous keratopathy that precluded pachymetry and specular microscopy.
A model of cultivated corneal endothelial transplantation for corneal endothelial dysfunction was established in primates whose corneal endothelial cells have less proliferative capacity in vivo. Our results suggest that this is a useful model for long-term observation in advance of the future clinical application of cultivated corneal endothelial transplantation.
探讨在灵长类动物模型中进行培养角膜内皮细胞移植的可行性。
从三只食蟹猴获取猴角膜内皮细胞(MCECs)进行培养,传代培养物在I型胶原载体上生长4周。刮除六只猴子右眼的角膜内皮,之后将培养的MCEC片植入四只眼的前房,并通过空气固定于Descemet膜。作为对照,将不含MCECs的胶原片移植到一只猴子的一只眼中,并将培养的MCECs悬液注入一只眼的前房。
培养的MCECs形成了紧密附着的六边形细胞汇合单层,显示出ZO-1和Na(+)-K(+)ATP酶表达。术后早期,MCEC片附着于Descemet膜,角膜透明度恢复。透明度恢复伴随着角膜厚度的降低。术后第7天在宿主角膜上检测到荧光素DiI标记的供体角膜内皮细胞。移植后6个月,MCEC移植的角膜保持透明,通过活体镜面显微镜观察到六边形细胞,密度为1992至2475个细胞/mm(2)。对照眼出现不可逆的大泡性角膜病变,无法进行测厚和镜面显微镜检查。
在体内角膜内皮细胞增殖能力较低的灵长类动物中建立了用于角膜内皮功能障碍的培养角膜内皮移植模型。我们的结果表明,这是一个在培养角膜内皮移植未来临床应用之前进行长期观察的有用模型。