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Abstract：
The aim of this study is to do a study of cryoinjury and ischaemic injury on testicular graft during cryopreservation and transplantation. According to time at 1, 3, 7 and 14 days after transplantation, the grafts were collected for immunohistochemistry assay for CD34 (blood vessel marker), VEGF (neoangiogenesis marker), caspase-3 (apoptosis marker) MAGE-A4 (germ cell marker). A significant increase was observed in the density of VEGF-positive blood vessels on day 3, reached a peak on day 7. On post-transplant day 3, a sharp increase occurred in the rate of spermatogonia-expressing caspase-3 until the day 7. At 14th day after transplantation, the spermatogonia number per round tubule of nonfrozen grafts was 41 ± 5.9% from that of fresh control tissues, while, in frozen-thawed grafts, the spermatogonia number per round tubule was 36.8 ± 4.6% from that of fresh control tissues. In testicular grafts, angiogenesis initiated reperfusion from day 3, and the formation of new blood vessel generally is completed about 7 days after transplantation. Angiogenesis in grafts after transplantation plays a crucial role in the restoration of function. Therefore, minimising ischaemic injury as well as improvement of cryopreservation protocols are needed to improve testicular graft after freezing, thawing and grafting.
摘要：
本研究的目的是研究冷冻保存和移植过程中睾丸移植物的冷冻损伤和缺血损伤。根据移植后1、3、7和14天的时间，收集移植物进行CD34（血管标志物）的免疫组织化学测定，VEGF（新血管生成标志物），caspase-3(凋亡标志物)MAGE-A4(生殖细胞标志物)。在第3天观察到VEGF阳性血管的密度显著增加,在第7天达到峰值。在移植后第3天，直到第7天，精原细胞表达caspase-3的比率急剧增加。移植后第14天,非冷冻移植物每根圆管的精原细胞数量为新鲜对照组织的41±5.9％，while,在冻融的移植物中，每个圆管的精原细胞数量为新鲜对照组织的36.8±4.6％。在睾丸移植物中，血管生成从第3天开始再灌注,并且新血管的形成通常在移植后约7天完成。移植后移植物中的血管生成在功能恢复中起着至关重要的作用。因此,最大限度地减少缺血损伤以及改善冷冻保存方案，以改善冷冻后的睾丸移植物，解冻和嫁接。
