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利用猪的器官为挽救脏器终末衰竭病人而进行的异种移植是有效救治病人的途径之一.由于猪细胞表面存在的α1,3半乳糖会引起超急性免疫排斥反应,导致移植器官最终被移植受体排斥.除人类和旧大陆猴外,所有哺乳动物细胞表面都有αGal半乳糖表达,但个体间表达量存在显著差异.为研究近交系五指山小型猪细胞表面α1,3半乳糖表达的个体差异,本实验利用FITC-isolectin进行荧光标记,通过流式细胞术以及激光共聚焦分析,对14例5月龄猪样本外周血单个核细胞表面的α1,3半乳糖水平进行检测.利用猪肾上皮PK15细胞确定FITC-isolectin标记的最适浓度为25ng/μl(细胞标记效率大于85%).结果显示,14头猪外周血单个核细胞表面α1,3半乳糖的表达差异范围在1.25~2.09倍之间,远低于普通非近交系猪的个体差异.本研究为利用近交系五指山小型猪作为异种器官移植的研究材料提供了基础数据.
Xenotransplantation using porcine organs to rescue patients with end-organ failure in the organ is one of the ways to effectively treat the patient.As the α1,3-galactose present on the surface of porcine cells causes hyperacute rejection, the transplanted organ is eventually replaced Transplant Receptor Rejection In all mammalian cells, human α-galactose and galactose expression were observed on the surface of all mammalian cells, but there was a significant difference between the expression of αGal and galactose In this experiment, FITC-isolectin was used for fluorescence labeling, and the α1,3-galactose levels on peripheral blood mononuclear cells of 14 piglets aged 5 months were detected by flow cytometry and confocal laser scanning. The optimal concentration of FITC-isolectin was 25ng / μl (cell labeling efficiency> 85%) in porcine renal epithelial PK15 cells.The results showed that the expression of α1,3-galactose on the surface of porcine renal epithelial cells was 1.25 ~ 2.09 times, which is much lower than that of normal non-inbred pigs.The present study provided the basis for the use of Wuzhishan minipig inbred line as a research material for xenotransplantation data.