兔角膜上皮细胞培养后增殖能力的观察

Experimental study of the proliferative ability of the cultivated rabbit corneal epithelial cells

目的:了解培养的兔角膜缘上皮细胞在原代及第2代细胞汇合时的增殖能力。方法:选健康新西兰白兔7只,显微镜下手术切取左眼上方少量角膜缘组织,采用组织块法分别进行培养,在原代上皮细胞汇合传代时(6鄄8d)和第2代细胞汇合时(10鄄12d),用流式细胞仪(FCM)检测样本的细胞周期和凋亡细胞。结果:培养的原代及第2代上皮细胞汇合时平均的增殖指数(PI)为19.1%和9.8%,凋亡细胞比例分别为0.57%和0.37%,处于增殖期细胞的比例在减少,而凋亡细胞的比例一直较低。结论:组织块法适合兔角膜缘上皮细胞培养,并且原代培养的兔角膜缘上皮细胞比第2代细胞具有更强的增殖能力,更适合进行细胞移植。

Objective: To study the proliferative ability of the cultivated rabbit corneal epithelial cells during the merging of cells of the primary generation and the second generation. Methods: A few limbal tissues at the upper part of left eyes were removed from seven selected healthy New Zealand white rabbits by operations under the microscope. The tissues were cultured and generated. The cell cycle and apoptosis cells of the samples were investigated by flow cytometry(FCM) during the merging of cells of the primary generation(6-8 d) and the second generation(10-12 d). Results: The average proliferative indexes during the merging of cells of the primary generation and the second generation were 19.1% and 9.8% respectively. The proportions of apoptosis cells were 0.57% and 0.37% respectively. The proportion of cells at the proliferative stage was decreasing, while the proportion of apoptosis cells remained small. Conclusion: The tissue-cultivating method is suitable for the culture of rabbit limbal epithelial cells. Moreover, the cultivated rabbit corneal epithelial cells of the primary generation are more active in proliferation than the second generation. Therefore, the former is more suitable for cell transplantation.