Urakawa Y, Miyazaki M, Ishihara A, Segawa K, Watanabe S, Shimizu Y
Department of Ophthalmology, Shinshu University School of Medicine, Matsumoto, Japan.
Jpn J Ophthalmol. 1988;32(4):401-11.
Human trabecular meshwork obtained from postmortem eyes was maintained in organ culture condition up to 12 weeks. The explants were dissected into specimens of three different sizes. Types 1 and 2 specimens were larger than 4 mm square and 1 mm thick, and type 3 specimens were about 1 mm3. Each specimen was placed in a depression made in the center of an agar block and the block was placed in a plastic culture dish. The space between the agar block and the culture dish was filled with liquid medium consisting of MEM Eagle supplemented with 10% fetal bovine serum, 10% chick embryo extract and 200 U/ml penicillin G. Afterwards, the explant was incubated in 5% CO2 with 100% humidity. Types 1 and 2 specimens maintained a normal appearance on electron microscopy during the entire 12 weeks of culture, while type 3 specimens showed marked cellular activation and cell loss soon after explantation. Morphometric studies regarding cell number in sections for light microscopy obtained from types 1 and 2 specimens revealed good preservation of trabecular cells during the 12 weeks of culture. Our culture system may be useful in investigating the direct effects produced on human trabecular meshwork by photocoagulation and various agents, for example anti-glaucoma drugs, steroids and viscoelastic materials.
从死后眼睛获取的人小梁网在器官培养条件下维持长达12周。将外植体切成三种不同大小的标本。1型和2型标本大于4平方毫米且厚1毫米,3型标本约为1立方毫米。每个标本置于琼脂块中央的凹陷处,然后将琼脂块放入塑料培养皿中。琼脂块与培养皿之间的空间填充有由添加了10%胎牛血清、10%鸡胚提取物和200 U/ml青霉素G的伊格尔MEM培养基组成的液体培养基。之后,将外植体在5%二氧化碳、湿度100%的条件下孵育。在整个12周的培养过程中,1型和2型标本在电子显微镜下保持正常外观,而3型标本在植入后不久就显示出明显的细胞活化和细胞丢失。对从1型和2型标本获取的用于光学显微镜检查的切片中的细胞数量进行的形态计量学研究表明,在12周的培养过程中小梁细胞保存良好。我们的培养系统可能有助于研究光凝和各种药物(例如抗青光眼药物、类固醇和粘弹性材料)对人小梁网产生的直接影响。