Laboratory of Biomedical Embryology, Università degli Studi di Milano, Milan, Italy.
Fertil Steril. 2013 Oct;100(4):1122-31. doi: 10.1016/j.fertnstert.2013.06.003. Epub 2013 Jul 3.
To compare conventional slow equilibrium cooling and directional freezing for cryopreservation of whole ovaries.
Experimental animal study.
Academic research environment.
ANIMAL(S): Adult ewes.
INTERVENTION(S): Eighty-one ovaries were randomly assigned to fresh control, conventional freezing (CF), and directional freezing (DF) group. Ovaries of CF and DF groups were perfused via the ovarian artery with Leibovitz L-15 medium, 10% fetal bovine serum, and 1.5 M dimethyl sulfoxide for 5 minutes. Each ovary was inserted into a glass test tube containing 10 mL of the same solution and cooled to -100°C or -70°C, respectively. Ovaries were stored in liquid nitrogen for a minimum of 2 weeks.
MAIN OUTCOME MEASURE(S): Structural integrity of cortical and medulla regions, vascular integrity, follicle in vitro development, cell proliferation, and DNA damage and repair.
RESULT(S): All examined parameters indicate that the structure of DF ovaries remains largely intact and comparable to fresh controls, whereas significant damages were observed in CF ovaries.
CONCLUSION(S): Directional freezing allows good preservation of whole ovaries, with most of the parameters taken into consideration almost identical to those recorded in fresh control samples. This encourages a reconsideration of the possible use of whole-ovary cryopreservation as a viable alternative to cortical fragments.
比较传统的缓慢平衡冷却法和定向冷冻法在保存整个卵巢中的作用。
实验动物研究。
学术研究环境。
成年母羊。
81 个卵巢被随机分配到新鲜对照组、传统冷冻（CF）组和定向冷冻（DF）组。CF 和 DF 组的卵巢通过卵巢动脉用 Leibovitz L-15 培养基、10%胎牛血清和 1.5M 二甲基亚砜灌注 5 分钟。每个卵巢被插入一个含有 10ml 相同溶液的玻璃试管中，并分别冷却至-100°C 或-70°C。卵巢在液氮中储存至少 2 周。
皮质和髓质区域的结构完整性、血管完整性、体外卵泡发育、细胞增殖以及 DNA 损伤和修复。
所有检查的参数都表明，DF 卵巢的结构基本保持完整，与新鲜对照组相当，而 CF 卵巢则观察到明显的损伤。
定向冷冻可以很好地保存整个卵巢，考虑到的大多数参数与新鲜对照组样本几乎相同。这鼓励重新考虑将整个卵巢冷冻保存作为皮质碎片的可行替代方案。
