System for bovine blastocyst production from lyophilized sperm and diploid cells

2015 Fiscal Year Final Research Report

• 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
• 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

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.

Free Research Field

応用動物科学