Department of Gynecology, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium.
Fertil Steril. 2011 May;95(6):2123.e9-12. doi: 10.1016/j.fertnstert.2011.01.014. Epub 2011 Feb 5.
To assess vitrification of prepubertal human testicular tissue in vitro.
Case report.
Academic research unit.
PATIENT(S): Two patients (6 and 12 years of age) who were to start gonadotoxic treatment for chronic granulomatous disease and acute lymphoblastic leukemia.
INTERVENTION(S): Long-term (10-day) organotypic culture performed immediately after vitrification and warming. Fresh tissue and tissue cryopreserved by slow-freezing were used as control samples.
MAIN OUTCOMES MEASURE(S): Spermatogonial cell survival (MAGE-A4) and proliferation (Ki67) were evaluated by immunohistochemistry (IHC) and tubular integrity by light microscopy.
RESULT(S): Qualitative analysis revealed that histologic characteristics of spermatogonia and Sertoli cells were preserved, as were cell-cell cohesion and cell adhesion to the basement membrane, in vitrified tissue as well as in frozen and fresh control samples. Survival of spermatogonia and their ability to proliferate as evidenced by IHC was also confirmed in cultured fresh, slow-frozen, and vitrified tissue.
CONCLUSION(S): Vitrification, having the advantage of being a faster and more convenient method, shows promise as an alternative strategy to slow-freezing in the emerging field of immature testicular tissue cryopreservation.
评估体外未成年人类睾丸组织的玻璃化。
病例报告。
学术研究单位。
两名患者（6 岁和 12 岁），因慢性肉芽肿病和急性淋巴细胞白血病需开始性腺毒性治疗。
玻璃化和复温后立即进行长期（10 天）器官型培养。新鲜组织和慢速冻存的组织作为对照样本。
通过免疫组织化学（IHC）评估精原细胞的存活（MAGE-A4）和增殖（Ki67），并通过光镜评估小管完整性。
定性分析显示，玻璃化组织以及冷冻和新鲜对照样本中均保留了精原细胞和支持细胞的组织学特征，细胞-细胞间的黏附力以及细胞与基底膜的黏附力也得以维持。IHC 也证实了培养的新鲜、慢速冻存和玻璃化组织中精原细胞的存活及其增殖能力。
玻璃化具有更快、更方便的优点，有望成为不成熟睾丸组织冷冻保存领域中替代慢速冻存的策略。
