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Effects of a gel culture system made of polysaccharides (xanthan gum and locust bean gum) on in vitro bovine oocyte development and gene expression of the granulosa cells
Molecular Reproduction and Development ( IF 2.5 ) Pub Date : 2021-06-06 , DOI: 10.1002/mrd.23518
Ayaka Sugimoto 1 , Yuki Inoue 1 , Keisuke Tanaka 2 , Akihisa Sinozawa 2 , Koumei Shirasuna 1 , Hisataka Iwata 1
Affiliation  

Xanthan gum (XG) and locust bean gum (LBG) are nontoxic polysaccharides that produce culture substrates. The present study examined the effect of XG-LBG gel on in vitro bovine oocyte growth and gene expression in granulosa cells. Oocytes and granulosa cell complexes (OGCs) were cultured in vitro on plastic culture plate (Plate) or XG-LBG gel for 16 days. OGCs formed a dome-like cavity surrounding the oocytes on plate but formed a spherical follicle structure on XG-LBG gel. The total granulosa cell numbers of the OGCs and their survival rate was greater for OGCs cultured on XG-LBG gel than for those cultured on plate. Oocytes grown on XG-LBG gels had higher lipid and mitochondrial content, as well as a larger diameter, than their plate counterparts. When oocytes grown in vitro were subjected to in vitro maturation and fertilization, the normal fertilization rate was significantly higher for oocytes developed on XG-LBG gel than that of oocytes cultured on the plate counterpart. RNAseq of the granulosa cells revealed that genes associated with focal adhesion, phosphatidylinositol 3'-kinase-Akt and Hippo signaling, and regulation of actin cytoskeleton were upregulated in granulosa cells of OGCs cultured on XG-LBG gel compared with those cultured on plate.

中文翻译:

多糖(黄原胶和槐豆胶)凝胶培养体系对体外牛卵母细胞发育和颗粒细胞基因表达的影响

黄原胶 (XG) 和刺槐豆胶 (LBG) 是产生培养基质的无毒多糖。本研究检测了 XG-LBG 凝胶对体外牛卵母细胞生长和颗粒细胞基因表达的影响。卵母细胞和颗粒细胞复合物 (OGC) 在塑料培养板 (Plate) 或 XG-LBG 凝胶上体外培养 16 天。OGC在板上形成围绕卵母细胞的圆顶状空腔,但在XG-LBG凝胶上形成球形滤泡结构。在 XG-LBG 凝胶上培养的 OGC 的总颗粒细胞数量及其存活率高于在平板上培养的 OGC。在 XG-LBG 凝胶上生长的卵母细胞比其平板对应物具有更高的脂质和线粒体含量,以及更大的直径。当体外生长的卵母细胞进行体外成熟和受精时,在 XG-LBG 凝胶上发育的卵母细胞的正常受精率明显高于在平板对应物上培养的卵母细胞。颗粒细胞的 RNAseq 显示,与平板培养的 OGC 颗粒细胞相比,在 XG-LBG 凝胶上培养的颗粒细胞中,与粘着斑、磷脂酰肌醇 3'-激酶-Akt 和 Hippo 信号传导以及肌动蛋白细胞骨架调节相关的基因上调。
更新日期:2021-07-24
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