人角膜上皮细胞的体外培养方法探讨

目的:探寻角膜上皮移植细胞的体外培养方法。方法:用改良的细胞悬液培养法与组织块培养法,描绘两种方法获取细胞的生长曲线,并计算细胞倍增时间;用~3H-胸腺嘧啶核苷(~3H-TdR)掺入与液体闪烁技术观察细胞DNA合成能力。结果:改良的细胞悬液法与组织块培养法培养的细胞平均倍增时间分别为54.15± 4.28小时和67.68±1.96小时(P<0.01);应用前者培养的细胞DNA合成也明显活跃(P<0.01)。结论:从角膜缘取材,应用改良的细胞悬液培养法收集的角膜缘上皮细胞含有角膜上皮干细胞,具有更强的分裂增殖能力,更适于体外培养的角膜上皮细胞移植之用。也证实了角膜上皮干细胞存在于角膜缘基底层的观点。眼科学报 1997;13:67～69。

Culture of Human Corneal Epithelial Cells

PURPOSE: To select culture technique of human epithelial cells for grafts. METHODS: Epithelial cells of limbus were collected by improved enzymatic disaggregation or explant technique growth curves and to calculate doubling time. DNA synthesis of cells were measured with 3H-thymidine incorporation and liquid scintillation techniques. RESULTS: Doubling time of cells which were collected by enzymatic disaggregation and by explant technique were 54.15 +/- 4.28 h and 67.88 +/- 1.96 h (P < 0.01). Cellular DNA synthesis of the former was more active (P < 0.01). CONCLUSIONS: Epithelial cells which were collected by improved enzymatic disaggregation from limbus include stem cells of corneal epithelium. These cells show more active cellulde proliferation and are fitter for use as grafts.