Development of polysaccharide gel culture system of in vitro embryo culture system.

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K22372
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 42:Veterinary medical science, animal science, and related fields
• Research Institution: Tokyo University of Agriculture
• Principal Investigator: Iwata Hisataka 東京農業大学, 農学部, 教授 (50385499)
• Project Period (FY): 2019-06-28 – 2022-03-31
• Keywords: 多糖ゲル / 培養基質 / 体外受精 / 体外発育卵子 / アクチン重合 / 胚盤胞

Outline of Final Research Achievements

The present study examined the effect of soft polysaccharides gel made of Xanthan gum (XG) and locust bean gum (LBG) on development of bovine oocytes and embryos. Oocytes-granulosa cells complexes (OGCs) derived from early antral follicles, oocytes derived from antral follicles and early developmental stage embryos were cultured on the XG-LBG gel. Using the XG-LBG gel as a culture substrate for in vitro oocyte growth, in vitro oocyte maturation, and in vitro development of embryos improved their developmental ability compared with those plastic plate-counterparts. RNA-Seq of the granulosa cells of OGCs showed that the differential expressed genes were associated with focal adhesion, phosphatidylinositol 3'-kinase-Akt and Hippo signaling, and regulation of actin cytoskeleton. Oocytes and embryos developed on the XG-LBG gel had higher expression levels of F-actin compared with those developed on the plate. LBG-XG gel is useful substrate for in vitro development of oocytes and embryos.

Free Research Field

胚発生

Academic Significance and Societal Importance of the Research Achievements

卵子や胚の発育にはシャーレ上のドロップが用いられる。プラスチックの特性が細胞の形態や分化に影響することが明らかにされつつあるが、本研究では卵子も胚も多糖ゲルで作成した基質の特性に反応して、その発育率や質を大きく改善することが明らかになった。多糖ゲル上では細胞内のmiRNAのプロファイルも大きく変化することが明らかになった。現行の培養環境は、基質性状から改善の余地が大きいことが明らかになった。