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Cryopreservation of oocytes and embryos by inducing the expression of cryoprotectant channels.

通过诱导冷冻保护剂通道的表达来冷冻保存卵母细胞和胚胎。

基本信息

批准号：
20380155
负责人：
EDASHIGE Keisuke
金额：
$ 11.98万
依托单位：
Kochi University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2008
资助国家：
日本
起止时间：
2008 至 2010
项目状态：
已结题

项目摘要

We tried to develop a vitrification method by which mouse oocytes and embryos at various stages can be cryopreserved by a single method. First, we examined whether the membrane-permeability of mouse oocytes and embryos can be equalized, because the permeability largely affects the condition for vitrification. We tried to induce the expression of water/cryoprotectant channels (AQP) in oocytes to increase the low membrane-permeability. We also tried in morulae to decrease the permeability of AQP3 which is markedly expressed in morulae and blastocysts. The cryoprotectant-permeability of oocytes increased slightly by retinoic acid but the maturation rate decreased remarkably. It was reported that the permeability of AQP3 decreased markedly at low pH. However, the permeability to ethylene glycol of mouse morulae, which expressed AQP3 abundantly, increased at low pH. Therefore, we considered that it would be difficult to make the membrane-permeability of oocytes/embryos equal. Next, we tried to develop a new vitrification solution in which a high concentration of sucrose was included to reduce the effect of membrane-permeability of oocytes/embryos on the vitrification condition. With this solution, embryos from the 2-cell to morula stages were successfully vitrified in the same condition, and vitrified embryos could be transported with dry ice (at -79℃). The method would be useful for cryopreservation of mouse embryos at various stages.

我们尝试开发一种玻璃化冷冻方法，使不同阶段的小鼠卵母细胞和胚胎可以通过一种方法进行低温保存。首先，我们检查了小鼠卵母细胞和胚胎的膜通透性是否可以均衡，因为通透性在很大程度上影响玻璃化的条件。我们尝试在卵母细胞中诱导水/冷冻保护剂通道（AQP）的表达以增加低膜通透性。我们还尝试在桑葚胚中降低AQP3的通透性，AQP3在桑葚胚和囊胚中表达明显。维甲酸可使卵母细胞的冷冻保护剂通透性略有提高，但成熟率明显降低。据报道，AQP3的通透性在低ph下明显降低，而表达AQP3丰富的小鼠桑皮对乙二醇的通透性在低ph下增加。因此，我们认为很难使卵母细胞/胚胎的膜通透性相等。接下来，我们尝试开发一种新的玻璃化液，其中含有高浓度的蔗糖，以降低卵母细胞/胚胎的膜通透性对玻璃化条件的影响。用该溶液在相同条件下成功地将2细胞胚玻璃化至桑葚胚，玻璃化后的胚可以用干冰（-79℃）运输。该方法可用于不同阶段小鼠胚胎的低温保存。

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Maintenance of fertility in cryopreserved Indian gerbil (Tateraindica)spermatozoa.

冷冻保存的印度沙鼠（Tateraindica）精子的生育能力的维持。

DOI：
发表时间：
2009
期刊：
Cryobiology 58(3)
影响因子：
0
作者：
C. Koshimoto;D. Watanabe;A. Shinohara;T. Morita
通讯作者：
T. Morita

ブタの卵子と拡大胚盤胞における水と耐凍剤の透過経路

猪卵母细胞和扩张囊胚中的水和冷冻保护剂渗透途径

DOI：
发表时间：
2009
期刊：
影响因子：
0
作者：
金波;東山龍一;米澤潤一;高橋誠一;三宅正史;葛西孫三郎;枝重圭祐
通讯作者：
枝重圭祐

Short-term storage of vitrified two一cell mouse embryos at-80°C for handy transportation with dry ice.

在 -80°C 下短期储存玻璃化两细胞小鼠胚胎，以便用干冰方便运输。

DOI：
发表时间：
2008
期刊：
影响因子：
0
作者：
Bo Jin;Eri Hotta;Yukiko Kobayashi;Shinsuke Seki;Masahiro Yoshimura;Hiroshi Honda;Keiji;Mochida;Atsuo Ogura;Magosaburo Kasai;Keisuke Edashige
通讯作者：
Keisuke Edashige

Handy transportation of vitrified mouse embryos with dry ice.

使用干冰方便运输玻璃化小鼠胚胎。