豚鼠血迷路屏障通透性的体外模型

Model in vitro for the permeability of blood labyrinth barrier from guinea pigs

目的 研究豚鼠血迷路屏障通透性体外模型的建立方法及其通透性特征 .方法 1用组织块贴壁培养法分离培养豚鼠耳蜗微血管内皮细胞 ,免疫组化法进行鉴定 ;2用小池技术建立起内皮细胞通透性的体外模型 ,并研究该模型中耳蜗、脑及肺 3种内皮细胞对 1 2 5I-牛血清白蛋白的通透性 ;3研究豚鼠血迷路屏障对 1 2 5I-牛血清白蛋白的通透性 .结果 1培养细胞致密融合时具有内皮细胞培养时典型的“铺路石样”外观 ,经免疫组化检测其内皮细胞标志性抗原 因子 ,95 %以上的培养细胞的胞质中呈棕黄色阳性反应 ;2耳蜗、脑及肺 3种内皮细胞的体外模型中加入 1 2 5I-牛血清白蛋白后 ,贝克 -时间变化图均呈抛物线型上升曲线 ,90 min内几乎呈直线 ;3豚鼠血迷路屏障对 1 2 5I-牛血清白蛋白的通透性曲线与体外模型相似 .

AIM To study the method of setting up model in vitro for the permeability of blood labyrinth barrier fromguinea pigs and the model's permeability. METHODS ① Cochlear microvascular endothelial cells (CMEC) from guinea pigs were isolated and cultured by tissue nubbles cultivation and were identified with immunohistochemical test, and the model in vitro for the permeability of CMEC was established using small pool technique; ② Researched the model's permeability on radioiodinated 125 I bovine serum album (RISA); ③ Studied the permeability on RISA of blood labyrinth barrier from guinea pigs in vivo . RESULTS ① The monolayer confluent cells showed typical 'flagstone' appearance as cultured endothelial cells, according to immunohistochemical detection of factor VIII, which is the mark antigen of endothelial cell, over 95% cells were observed the positive reaction with cytoplasmic buffy pigsmenting; ② After RISA administration, the Bake time curve of the model took on an ascending parabola, while within 90 min appeared like a straight line; ③ Permeability curve on RISA of the model in vitro simillar to that of blood labyrinth barrier in vivo . CONCLUSION The model in vitro of blood labyrinth barrier reveals the permeability of in vivo .