System for bovine blastocyst production from lyophilized sperm and diploid cells
Project/Area Number |
24580407
|
Research Category |
Grant-in-Aid for Scientific Research (C)
|
Allocation Type | Multi-year Fund |
Section | 一般 |
Research Field |
Applied animal science
|
Research Institution | Shinshu University |
Principal Investigator |
HOCHI Shinichi 信州大学, 学術研究院繊維学系, 教授 (10283243)
|
Project Period (FY) |
2012-04-01 – 2016-03-31
|
Project Status |
Completed (Fiscal Year 2015)
|
Budget Amount *help |
¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2012: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
|
Keywords | 凍結乾燥 / コラプス / α-ヘモリジン / トレハロース / ROCK阻害剤 / ビタミンE / 体細胞クローン / ICSI / SCNT / ケーキ / ウシ精子 / Tg' / 含水率 / 胚盤胞発生率 / αヘモリジン / ウシ顆粒膜細胞 / ケーキコラプス / DNA損傷 / アルカリコメットアッセイ / クローン胚盤胞 / フリーズドライ |
Outline of Final Research Achievements |
The aim of this study was to establish blastocyst production system from freeze-dried (FD) sperm and diploid cells in the bovine species. Results obtained were: (1) Incorporation of trehalose using alpha-hemolysin mutant did not inhibit DNA damage in FD granulosa cells. (2) Incidence of FD cake collapse was related with the glass transition temperature of the maximally freeze-concentrated phase (Tg'), and trehalose in FD buffer contributed to increase the Tg' and to inhibit the cake collapse. (3) ROCK inhibitor or alpha-tocopherol supplemented to recovery culture medium could rescue metaphase-II oocytes after vitrification and warming. (4) Morphologically normal blastocysts derived from both FD spermatozoa (via intracytoplasmic sperm injection) and FD fibroblasts (via somatic cell nuclear transplantation) were successfully produced in vitro.
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Report
(5 results)
Research Products
(28 results)