Department of Gynecology, Medical School, Institute of Experimental and Clinical Research, Université catholique de Louvain, 1200 Brussels, Belgium.
Theriogenology. 2012 Mar 15;77(5):1008-13. doi: 10.1016/j.theriogenology.2011.10.015. Epub 2011 Dec 6.
This study demonstrates preservation of tissue integrity, maintenance of proliferating spermatogonia and Leydig cell functionality after vitrification and transplantation of non-human primate immature testicular tissue. The objective was to assess the potential of vitrification of non-human primate immature testicular tissue (ITT) in an in vivo xenotransplantation model. Testicular tissue was obtained from one immature rhesus monkey (Macaca mulatta) aged 4 years. Collection and vitrification of testicular tissue, followed by short-term xenografting (3 wks) to nude mice were performed to evaluate and compare vitrified/warmed and fresh tissue. Fresh ungrafted tissue was used for control purposes. Cell density and seminiferous tubule (ST) integrity were assessed by light microscopy. Presence of spermatogonia (SG) (MAGE-A4), proliferation (Ki-67) and Leydig cell (LC) functionality (3β-hydroxysteroid dehydrogenase; 3β-HSD) were evaluated by immunohistochemistry (IHC). Qualitative analysis revealed preservation of the histologic characteristics of SG and Sertoli cells (SCs), as well as cell-cell cohesion and cell adhesion to the basement membrane, in both vitrified and fresh grafted tissues. Survival of SG able to proliferate and functional LCs was confirmed by IHC in fresh and vitrified grafts. In conclusion, vitrification appears to be a promising approach, representing an alternative strategy to slow-freezing in the emerging field of ITT cryopreservation and cryobanking.
本研究证明了在非人类灵长类动物未成熟睾丸组织（ITT）的体内异种移植模型中，玻璃化和移植后组织完整性得以保留，增殖性精原细胞和莱迪希细胞（Leydig cell）功能得以维持。本研究的目的是评估非人类灵长类动物未成熟睾丸组织（ITT）玻璃化的潜力。从一只 4 岁的幼年恒河猴（Macaca mulatta）中获取睾丸组织。进行睾丸组织的采集和玻璃化，然后进行短期异种移植（3 周）至裸鼠，以评估和比较玻璃化/加热和新鲜组织。未移植的新鲜组织用于对照目的。通过光镜评估细胞密度和生精小管（ST）完整性。通过免疫组织化学（IHC）评估精原细胞（SG）（MAGE-A4）、增殖（Ki-67）和莱迪希细胞（LC）功能（3β-羟类固醇脱氢酶；3β-HSD）。定性分析显示，在玻璃化和新鲜移植组织中，SG 和支持细胞（SCs）的组织学特征、细胞-细胞黏附以及细胞与基底膜的黏附得以保留。在新鲜和玻璃化移植组织中，通过 IHC 证实了能够增殖和具有功能的 LC 的 SG 的存活。总之，玻璃化似乎是一种很有前途的方法，代表了 ITT 冷冻保存和低温保存领域中慢速冻融的替代策略。
