Baudot A, Courbiere B, Odagescu V, Salle B, Mazoyer C, Massardier J, Lornage J
Département MCBT, Institut Néel - CNRS/UJF, BP 166, 38042 Grenoble Cedex 09, France.
Cryobiology. 2007 Dec;55(3):236-48. doi: 10.1016/j.cryobiol.2007.08.004. Epub 2007 Aug 24.
Cryopreservation of ovarian tissue aims to assist young women who require treatments that may lead to sterility or infertility. Cryopreservation procedures should therefore be as simple and efficient as possible. This study investigates rapid cooling outcomes for whole sheep ovaries. Ovaries were perfused with VS4 via the ovarian artery, and cooled by quenching in liquid nitrogen in less than a minute (estimated cooling rate above 300 degrees C/min till the vitreous transition temperature). The ovaries were rewarmed in two stages: slow warming (12-16 degrees C/min from -196 to -133 degrees C) in liquid nitrogen vapour, followed by rapid thawing in a 45 degrees C water bath at about 200 degrees C/min. DSC measurements showed that under these cryopreservation conditions VS4 would vitrify, but that VS4 perfused ovarian cortex fragments did not vitrify, but formed ice (around 18.4%). Immediately following rewarming, a dye exclusion test indicated that 61.4+/-2.2% of small follicles were viable while histological analysis showed that 48+/-3.8% of the primordial follicles were normal. It remains to be clarified whether follicle survival rates will increase if conditions allowing complete tissue vitrification were used.
卵巢组织冷冻保存旨在帮助那些需要接受可能导致不育或不孕治疗的年轻女性。因此，冷冻保存程序应尽可能简单有效。本研究调查了整只绵羊卵巢的快速冷却结果。通过卵巢动脉用VS4灌注卵巢，并在不到一分钟的时间内于液氮中骤冷（估计冷却速率高于300℃/分钟直至玻璃化转变温度）。卵巢分两个阶段复温：在液氮蒸气中缓慢升温（从-196℃至-133℃，升温速率为12 - 16℃/分钟），随后在45℃水浴中以约200℃/分钟的速率快速解冻。差示扫描量热法测量表明，在这些冷冻保存条件下，VS4会玻璃化，但用VS4灌注的卵巢皮质碎片不会玻璃化，而是形成冰（约18.4%）。复温后立即进行的染料排除试验表明，61.4±2.2%的小卵泡存活，而组织学分析显示48±3.8%的原始卵泡正常。如果使用允许组织完全玻璃化的条件，卵泡存活率是否会提高仍有待阐明。
