Hasegawa Akiko, Mochida Nahoko, Ogasawara Toshitada, Koyama Koji
Laboratory of Developmental Biology and Reproduction, Institute for Advanced Medical Sciences, Nishinomiya, Japan.
Fertil Steril. 2006 Oct;86(4 Suppl):1182-92. doi: 10.1016/j.fertnstert.2005.12.082. Epub 2006 Sep 11.
To determine an optimum condition for vitrification of various animal species and to examine the developing capacity into pups of mouse oocytes in preantral follicles originating from cryopreserved ovaries.
Experimental animal study.
Laboratory environment.
ANIMAL(S): Normal (C57BL/6 x DBA2) F1 mice.
INTERVENTION(S): Vitrification of animal ovaries using polyester sheets as a storage device; collection of oocyte-granulosa cell complexes by enzymatic treatment; and in vitro growth (IVG), in vitro maturation (IVM), and IVF, with embryo transplantation to pseudopregnant mice.
MAIN OUTCOME MEASURE(S): Histological analysis of vitrified and warmed ovaries from several animal species; measurement of successful rates in IVG, IVM, and IVF of mouse oocytes in oocyte-granulosa cell complexes collected from the vitrified and warmed ovaries; and achievement of pup birth.
RESULT(S): The vitrification method used was effective for storage of various animal ovaries. The oocytes enclosed in preantral follicles that were yielded from the vitrified-warmed ovaries preserved the capacity for developing into pups after IVG, IVM, and IVF.
CONCLUSION(S): The findings indicated that ovarian cryopreservation by vitrification and subsequent IVG, IVM, and IVF are promising methods for restoring young cancer patients' fertility. The practical information presented here is applicable to human ovarian tissues.
确定不同动物物种玻璃化冷冻的最佳条件,并检测源自冷冻保存卵巢的窦前卵泡中小鼠卵母细胞发育成幼崽的能力。
实验动物研究。
实验室环境。
正常(C57BL/6×DBA2)F1小鼠。
使用聚酯片作为储存装置对动物卵巢进行玻璃化冷冻;通过酶处理收集卵母细胞-颗粒细胞复合体;进行体外生长(IVG)、体外成熟(IVM)和体外受精(IVF),并将胚胎移植到假孕小鼠体内。
对几种动物物种的玻璃化冷冻及复温后的卵巢进行组织学分析;测定从玻璃化冷冻及复温后的卵巢收集的卵母细胞-颗粒细胞复合体中,小鼠卵母细胞IVG、IVM和IVF的成功率;以及幼崽出生情况。
所采用的玻璃化冷冻方法对多种动物卵巢的保存有效。从玻璃化冷冻及复温后的卵巢中获得的窦前卵泡内的卵母细胞,在经过IVG、IVM和IVF后仍保留发育成幼崽的能力。
研究结果表明,通过玻璃化冷冻保存卵巢以及随后的IVG、IVM和IVF是恢复年轻癌症患者生育能力有前景的方法。此处提供的实用信息适用于人类卵巢组织。