Bovine eight cell stage embryos derived from oocytes matured on plastic dish or Xanthan and Locust been gum gel
In vivo oocyte maturation is conducted on soft matrix consist of granulosa cells mass in follicles, whereas in vitro oocyte maturation is conducted on rigid plastic plate. It has been reported that soft polysaccharide gel made of Xanthan and Locust been gum significantly stimulated the growth of the bovine oocytes and development of embryos (Mol Reprod Dev. 2021;88:516-524, Reprod Domest Anim. 2020 55:1124-1131). This data is RNAseq of eight cell stage embryos 48h after insemination. Ovaries were collected at a slaughterhouse and transported to the laboratory within 4 hours. Oocytes were collected from bovine antral follicles (3-5 mm in diameter) of Japanese Black Cows and cultured in TCM-199 medium containing 10ng/ml EGF and 10% FCS. At the end of maturation period, the oocytes were fertilized and incubated for 18h. And then presumptive zygotes were denuded and incubated in synthetic oviductal fluid medium containing 1% FCS for 30h. Eight stage embryos were subjected to RNA seq. RNA extraction was conducted using RNAqueous (Invitrogen). The RNA quality and concentration were examined using a Bioanalyzer (Agilent technologies, Palo Alto, CA, USA). cDNA of the oocytes was produced from each 150pg RNA using SMART Seq v4 UltraÂ® Low Input RNA Kit for Sequencing Clontech). Library was constructed using NEBNext Single Cell/Low Input RNA Library Prep Kit The quality and quantity were determined using the Agilent 2100 Bioanalyzer, followed by re-measurement using Kapa Library Quantification Kit Kapa Biosystems. and sequencing was conducted using NextSeq1000 Single read 1 x 100 bp. Image analysis, base calling and quality filtering were performed using the RTA version 2.4.11 (Illumina) following the manufacturer's instructions, and the sequence data was converted to Fastq using bcl2fastq2 v2.20.0.422. Sequence data were filtered to discard the adapter sequence, ambiguous nucleotides and low-quality sequences.