miRNA in granulosa cells cultured under differential conditions

Follicular fluid contained miRNAs at a high concentration and the miRNA play a crucial role in oocyte growth. These miRNAs are believed to be secreted from granulosa cells (GCs). The data are miRNA extracted from the GCs. Oocyte-granulosa cells complexes (OGCs) were collected from early antral follicles (0.5-0.7mm in diameter) on the surface of ovaries. The OGCs were cultured on plastic plate (96 well culture plate) or on gels made of polysaccharides (1% Xanthan gum and Locust bean gum) for 16 days. The gel culture condition improved oocyte development in cows and pigs, which is reported in PMID: 34096128 PMID: 31588063. At day 16 of in vitro culture, GCs were collected from well-developed OGCs. RNA in the GCs was extracted using Nucleospin miRNA kit (740971.50, Takara, Kusatsu, Japan). RNA quality and concentration were examined using a Bioanalyzer (Agilent technologies, Palo Alto, CA, USA). cDNA library was constructed using SMARTer miRNA Seq Kit for Illumina (TaKaRa). The multiplied samples were sequenced as 75 bp reads (single read) on the NextSeq 500 (Illumina). Image analysis, base calling and quality filtering were performed using the bcl2fastq2 v2.18.0.12 (Illumina) following the manufacture's instructions.