Biophysical Society Thematic Meeting | Singapore

Mechanobiology of Disease

Poster Abstracts

3-POS Board 3 Comprehensive Analyses of Gene Expression Patterns in Early Mouse Early Embryos

Experiencing Chemical and Mechanical Stimuli Yuka Asano 1,2 , Koji Matsuura 1,2 .Keiji Naruse 2 .

2 Okayama University, Okayama, Japan. 1 Okayama University of Science, Okayama, Japan, In vitro mammalian embryonic development can be enhanced by mimicking physiological conditions, which can provide better infertility treatment. By comprehensive analyses of gene expression, we evaluated differences in gene expression patterns of ICR mouse blastocysts cultured in vitro from two-cell embryo stage using different media under a static or dynamic culture system to subject the embryos to mechanical stimuli (MS). To compare the influences of the medium on early mouse embryonic development, we used mW medium (mW-S) and KSOM medium (KSOM-S) under static culture. The influences of MS in mW medium were investigated using the tilting embryo culture system (TECS), which improves blastocyst development. We compared the gene expression patterns between the TECS (mW-T) and mW-S groups. Blastocyst development rates in the mW-S, KSOM-S, and mW-T groups were 62.6%, 94.1%, and 70.8%, respectively (P < 0.01). We extracted total RNAs thrice from the three groups, and the total number of blastocysts was approximately 80 × 9 = 720. RNA amplification, RNA-Seq library preparation, and RNA sequencing were consigned to Hokkaido System Science Co. Ltd. Upon comparison of the relative gene expression levels between the KSOM-S and mW-S groups, the gene expression level in the trophectoderm (TE), which functions as a source of embryonic nutrition, was improved in culture conditions under KSOM medium compared with that in culture conditions under mW medium. Tagln2 gene expression as a TE marker increased (P < 0.01) under KSOM culture. Gdf3 gene expression levels related to epiblast (EPI) development in the mW-T group was higher than that in the mW-S group (P < 0.01). MS improved EPI development under the same medium conditions. The expression patterns of EPI and TE markers differed between the culture systems with chemical stimuli by medium and MS. We consider that optimized medium components can improve both TE and EPI development and that MS can improve the gene expression of EPI markers. However, the gene expression level induced by MS would be relatively lower in TE than in EPI.

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