Eye Clinic, DiNOGMI, University of Genoa and IRCCS San Martino Polyclinic Hospital, Genoa, Italy
Laboratory of Regenerative Medicine, Department of Oncology, Biology and Genetics, IRCCS San Martino Polyclinic Hospital, Genoa, Italy.
In Vivo. 2019 Nov-Dec;33(6):1851-1855. doi: 10.21873/invivo.11678.
To develop a method capable of identifying human corneal limbal stem cells (LSCs) and follow their proliferation and migration in the epithelium.
Ten fresh matched pairs of cadaveric normal human corneas were obtained from donors. Carboxyfluorescein diacetate succinimidyl ester (CFSE) was used to target LSCs. The distribution of CFSE-positive cell clusters was analyzed by fluorescence microscopy by counterstaining with 4',6-diamidino-2-phenylindole (DAPI). Fluorescence was digitally recorded for seven days, and the rate of cell movement was determined.
CFSE-labeled cells were tracked in corneas. Analysis of time sequences revealed that they moved centripetally. Daily average CFSE-labeled LSC movement was 0.073±0.01 cm (±SD).
CFSE allowed us to identify LSCs and to track their centripetal migration from the limbal basal layer to the anterior ocular surface. This experimental system appears to be a valuable tool for further studies on corneal epithelial cell migration and proliferation.
开发一种能够识别人眼角膜缘干细胞（LSCs）并追踪其在眼表上皮细胞中增殖和迁移的方法。
从供体中获得 10 对新鲜匹配的正常人体角膜。使用羧基荧光素二乙酸琥珀酰亚胺酯（CFSE）标记 LSCs。通过用 4'，6-二脒基-2-苯基吲哚（DAPI）复染，荧光显微镜分析 CFSE 阳性细胞簇的分布。连续 7 天对荧光进行数字化记录，并测定细胞运动速度。
CFSE 标记的细胞在角膜中被追踪。时间序列分析表明，它们向心性移动。每日平均 CFSE 标记的 LSC 运动速度为 0.073±0.01cm（±SD）。
CFSE 使我们能够识别 LSCs，并追踪其从角膜缘基底层向眼前部表面的向心性迁移。该实验系统似乎是研究角膜上皮细胞迁移和增殖的有用工具。
