The developmental ability of vitrified oocytes from different mouse strains assessed by parthenogenetic activation and intracytoplasmic sperm injection
The developmental ability of vitrified oocytes from different mouse strains assessed by parthenogenetic activation and intracytoplasmic sperm injection
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DOI：
10.1262/jrd.19058
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发表时间：
2007-12-01
影响因子：
1.8
通讯作者：
Ogura, Atsuo
中科院分区：
文献类型：
--
作者：
Endoh, Keisuke;Mochida, Keiji;Ogura, Atsuo
评价卵母细胞冻融后的发育能力是优化卵母细胞冷冻保存技术的重要环节。然而，小鼠卵母细胞冻融后的体外受精往往是低效的，因为精子在缺乏周围卵丘细胞的情况下不能完全获能。本研究旨在通过孤雌激活和卵胞浆内单精子注射（ICSI）后卵母细胞的发育情况来评价不同品系小鼠卵母细胞的冷冻保存效果。从杂交（C57 BL/6 x DBA/2）F1或近交（C57 BL/6 J、C3 H/HeN、DBA/2 J和BALB/cA）品系收集卵母细胞，并在含有乙二醇、DMSO、Ficoll和蔗糖的溶液中玻璃化冷冻。在第一系列的实验中，卵母细胞被激活的Sr 2+处理后，温暖的孤雌生殖。从近交系的卵母细胞，但不是那些的F1杂交，二倍体细胞松弛素处理，以获得足够数量的囊胚。在所有测试的菌株中，来自玻璃化卵母细胞的孤雌胚胎发育成囊胚的比率在23%至68%之间。在第二系列实验中，将来自每个菌株的玻璃化冷冻卵母细胞在加温后注射同源精子。从所有品系中获得正常后代，每个胚胎移植率为5 - 26%。因此，卵母细胞冷冻保存方案的可行性可以通过孤雌胚胎的体外发育或ICSI胚胎的体内发育来容易地评估。此外，这四种主要近交系小鼠的卵母细胞可以安全地冷冻保存用于生产后代。
Assessment of the developmental ability of oocytes following freezing and thawing is an important step for optimizing oocyte cryopreservation techniques. However, the in vitro fertilization of frozen-thawed mouse oocytes is often inefficient because of incomplete capacitation of spermatozoa in the absence of surrounding cumulus cells. This study was undertaken to determine whether the oocyte cryopreservation efficiency of different strains of mice could be assessed from the development of oocytes following parthenogenetic activation and intracytoplasmic sperm injection (ICSI). Oocytes were collected from hybrid (C57BL/6 x DBA/2) F1 or inbred (C57BL/6J, C3H/HeN, DBA/2J and BALB/cA) strains and were vitrified in a solution containing ethylene glycol, DMSO, Ficoll and sucrose. In the first series of experiments, oocytes were activated parthenogenetically by Sr2+ treatment after warming. The oocytes from the inbred strains, but not those of the F1 hybrid, were diploidized by cytochalasin treatment to obtain a sufficient number of blastocysts. In all strains tested, parthenogenetic embryos derived from vitrified oocytes developed into blastocysts at rates between 23 and 68%. In the second series of experiments, vitrified oocytes from each strain were injected with homologous spermatozoa after warming. Normal offspring were obtained from all strains at rates between 5 and 26% per embryo transferred. Thus, the feasibility of oocyte cryopreservation protocols can be assessed easily by in vitro development of parthenogenetic embryos or by in vivo development of ICSI embryos. Moreover, the oocytes of these four major inbred strains of mice can be cryopreserved safely for production of offspring.