| Project/Area Number |
20380155
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Applied animal science
|
|---|
| Research Institution | Kochi University |
|---|
Principal Investigator |
EDASHIGE Keisuke Kochi University, 教育研究部・総合科学系, 教授 (30175228)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
KASAI Magosaburo 高知大学, 教育研究部・総合科学系, 教授 (60152617)
KOSHIMOTO Chihiro 宮崎大学, フロンティア科学実験総合センター, 教授 (70295210)
|
|---|
| Project Period (FY) |
2008 – 2010
|
| Project Status |
Completed (Fiscal Year 2010)
|
| Budget Amount *help |
¥18,720,000 (Direct Cost: ¥14,400,000、Indirect Cost: ¥4,320,000)
Fiscal Year 2010: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2009: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2008: ¥11,050,000 (Direct Cost: ¥8,500,000、Indirect Cost: ¥2,550,000)
|
|---|
| Keywords | 凍結保存 / 卵子 / 胚 / 水透過性 / 耐凍剤透過性 / アクアポリン / 受精卵 / 促進拡散 |
|---|
| Research Abstract |
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.
|
