Rahimi G, Isachenko V, Todorov P, Tawadros S, Mallmann P, Nawaroth F, Isachenko E
Department of Obstetrics and Gynaecology, University of Cologne, Cologne, Germany.
Cryo Letters. 2009 Jul-Aug;30(4):300-9.
One of the new emerging techniques to preserve reproductive potential of cancer patients is cryopreservation of ovarian fragments prior to medical treatment and their retransplantation after healing. In order to investigate and compare apoptosis in human ovarian tissue after conventional ("slow") freezing and vitrification, we used a xenograft model in which conventionally frozen, vitrified and fresh non treated human ovarian tissue pieces were subcutaneously transplanted in SCID mice. The tissue samples were weekly, during four weeks, recovered from scarified SCID mice. The apoptosis was examined by immunohistochemical staining with the anti-caspase-3 antibody. There was a significant difference between the amount of apoptotic cells in cryopreserved ovarian tissue independent from mode of cooling compare to the control. The ovarian tissue after vitrification showed a significantly higher amount of apoptotic cells, than in slow frozen. The results obtained after comparative study of two different cryopreservation methods show that vitrification of human ovarian tissue could become a practice-relevant alternative to slow cryopreservation only after further improvement.
保存癌症患者生殖潜能的新兴技术之一是在医疗治疗前冷冻保存卵巢组织片段,并在治愈后进行再移植。为了研究和比较传统(“慢速”)冷冻和玻璃化后人类卵巢组织中的细胞凋亡情况,我们使用了一种异种移植模型,将传统冷冻、玻璃化和新鲜未处理的人类卵巢组织块皮下移植到SCID小鼠体内。在四周内,每周从处死的SCID小鼠中取出组织样本。通过用抗半胱天冬酶-3抗体进行免疫组织化学染色来检测细胞凋亡。与对照组相比,冷冻保存的卵巢组织中凋亡细胞数量与冷却方式无关,存在显著差异。玻璃化后的卵巢组织显示出的凋亡细胞数量明显高于慢速冷冻的组织。对两种不同冷冻保存方法的比较研究结果表明,人类卵巢组织的玻璃化只有在进一步改进后才能成为与传统冷冻保存相关的实用替代方法。