目的:通过研究深低温保存前后大月龄胎儿主、肺动脉带瓣血管组织相容性白细胞抗原(HLA)的表达情况,了解HLA抗原的表达位置以及深低温保存过程对HLA抗原表达的影响。
方法:取脑死亡胎儿(胎龄28周以上)主、肺动脉带辦血管10例,热缺血时间要求在12小时之内。每例标本分成两部分,一部分新鲜者作为对照组;另一部分为实验组经M199原液与二甲基亚砜(DMSO)处理后浸入液氮中深低温冻存。所有标本用冰冻切片包埋剂(ZLI-9302)放在不锈钢包埋模具上快速冷冻。它们被储存在—20℃冰箱,然后用恒冷箱切片机(Leica CM 1900)切片。所有切片包在银箔中储存在—20℃冰箱中直到被染色,用免疫组化的方法利用鼠抗人单克隆抗体:HLA-ABC(0736)、HLA-DR(ZM-0136),CD45RO(ZM-0055),CD34(ZM-0046),检测冷冻前后人组织相容性白细胞抗原Ⅰ类(HLA ABC)、Ⅱ类(HLADR,DP,DQ)和内皮细胞的变化情况,并将检测结果作图像分析。
结果:新鲜的血管壁和瓣膜内皮表面均表达Ⅰ类抗原(+++)一(++++),而Ⅱ类抗原在血...
【英文摘要】
Objective: The present study was undertaken to analysis the HLA alloantigen expression on cardiac homografts.To evaluate the effect of cryopreservation on immunogenicity of the homografts aortic and pulmonary conduits and valves from late-pregnancy fetus.Methods: Aortic and pulmonary HVCs were procured from 3 male and 7 female brain death fetus (with gestational age more than 28 weeks,WIT within 12 hours).Every specimen cut into two portions,the fresh one portion was control group and were quickly frozen wi...