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为探讨“简化四步法”深低温冷冻保存角膜材料的角膜细胞活性,采用体外组织培养技术对深低温保存的兔角膜的上皮细胞、内皮细胞和基质细胞进行培养和观察。结果表明,上皮细胞一般在培养5～7天贴壁,形成单层,形态呈不规则形或多边形;2周以后逐渐出现老化和脱落现象,仅能传1代。内皮细胞2～3天可贴壁,形成单层,但未能显示六边形,仅为不规则形或圆形,不易传代。基质细胞贴壁时间在培养后1周,形成整齐和密度较高的单层,能传2～3代。结论:用“简化四步法”长期深低温冷冻保存的角膜材料细胞尚有良好的活性,证实可用于角膜移植。
In order to investigate the corneal cell activity of corneal cryopreservation cryopreservation by “Simplify Four Steps Method”, the epithelial cells, endothelial cells and stromal cells of rabbit cornea preserved in cryogenic temperature were cultured and observed by in vitro tissue culture. The results showed that the epithelial cells generally adherent to form a monolayer after 5 to 7 days of culture, with irregular or polygonal shape. After 2 weeks, the epithelial cells gradually appeared to be aged and shedding, which could only pass on to the first generation. Endothelial cells 2 to 3 days can be attached to the wall, the formation of a single layer, but failed to show hexagonal, irregular or circular only, not easy passage. Stromal cell adherent time in the culture after 1 week, forming a neat and high density of monolayers, can pass 2 to 3 generations. CONCLUSION: The corneal material cells cryopreserved by the simplified four-step method have good activity and have been proved to be suitable for corneal transplantation.