Granulosa cells derived from young and aged cows

Granulosa cells are important for oocyte growth. Aging reduces oocyte quality which is a causal factor for aged associated infertility. Ageing associated events in granulosa cells will help understanding the aged associated low quality oocytes. This data is RNAseq of the granulosa cells comes from antral follicles (3-5 mm in diameter) of young (25-50 month in age) and aged (over 140 month in age) Japanese Black Cows. Ovaries were collected at a slaughterhouse and transported to the laboratory within 4 hours. And then granulosa cells were aspirated from antral follicels. These granulosa cells were cultured in medium (TCM-199 medium containing 5% FCS) on shares and attached survived granulosa cells were collected after 24h of incubation. The granulosa cells were collected from shares and used for RNA seq. RNA in granulosa cells were extracted using RNAqueous (Invitrogen) and the RNA quality and concentration were examined using a Bioanalyzer (Agilent technologies, Palo Alto, CA, USA). Library quality and quantity were determined using the Agilent 2100 Bioanalyzer and the KAPA Library Quantification Kit (KAPA Biosystems, Wilmington, MA, USA), respectively. Clusters were generated on a cBot (Illumina), and one lane of the multiplied samples was sequenced as 75 bp reads (single read) on the NextSeq (Illumina). Image analysis, base calling and quality filtering were performed using the bcl2fastq2 v2.18.0.12 (Illumina) following the manufacturer's instructions. Sequence data were filtered to discard the adapter sequence, ambiguous nucleotides and low-quality sequences. The remaining sequence data were aligned to the Bos Taurus genome sequence (ARS-UCD1.2/bosTau9) to count sequence reads.