RNA expression of Cx37 and Cx43 in bovine oocytes and cumulus cells after in vitro maturation depending on the presence of cumulus cells in the culture system (fresh or after cryopreservaion)

This dataset has been presented in our paper "New Approach to the Cryopreservation of GV Oocytes andCumulus Cells through the Lens of Preserving the IntercellularGap Junctions Based on the Bovine Model" The dataset represents the RNA expression of Cx37 and Cx 43 in oocytes and cumulus cells depending on the presence of cumulus cells in the culture system (fresh or after cryopreservaion) The obtained COCs from bovine ovaries were divided into groups and matured in vitro in a dropof IVM medium (Stroebech, Copenhagen, Denmark) with a volume of 100 μL under mineraloil. Groups: fresh immature COCs (immCOCs), fresh COCs after IVM (fCOCs), cryopreserved COCs after IVM (cryo-COCs), freshly denuded oocytes after IVM (fDOs), cryopreserved denuded oocytes afterIVM (cryoDOs), fresh denuded oocyte matured in vitro with a fresh suspension of CCs(fDOs + fCCs), cryopreserved oocytes matured in vitro with a cryopreserved suspensionof CCs (cryoDOs + cryoCCs), fresh CCs after culturing in IVM media (fCCs), cryopreservedcumulus cells after culturing in IVM media (cryoCCs). COCs or DOs and CCs were subjected to RNA isolation using the PicoPure RNAIsolation Kit (#KIT0204, Applied Biosystems, Waltham, MA, USA) according to the manufacturer’sinstructions. The RNA samples were stored at −80 ◦C. The RNA content in eachsample was 60 ng. The resulting complementary DNA (cDNA) was stored at −20 ◦C forfurther quantitative PCR (qPCR) analysis. TaqMan Universal Master Mix II (Life Technologies)was used. For each of the genes (Table 1), the PCR mix consisted of 3 μL of cDNA(equal to 2 ng), 0.5 μL of the TaqMan assay, 5 μL of the TaqMan master mix, and 1.5 μLof RNase-free water. The mRNA transcript levels were normalized to GAPDH (internalcontrol). The 384-well plates were prepared using the Bravo automated liquid handlingplatform (Agilent Technologies, Inc., Santa Clara, CA, USA). The analysis was performedin duplicate for each sample, in a final volume of 10 μL, using the ABI Prism 7900 HTReal-Time PCR System. The thermal profile used to amplify the genes was as follows:polymerase activation (95 ◦C for 10 min), followed by 45 cycles of denaturation (95 ◦C for15 s) and annealing (60 ◦C for 1 min). Real-time PCR Miner Software 4.0 (118.190.66.83) [18]was used to estimate the mean efficiency of PCR amplification and the cycle threshold (Ct)values for each gene. We conducted at least three biological replicates for the expression of eachgene in each group.