GSM8819054: D21 8F mini-oocyte rep2; Mus musculus; RNA-Seq
4F- or 8F-mESCs routinely cultured were detached from culture plates by 1× TrypLE solution (Thermo), and the detached cells were thoroughly dissociated into single cells by pipetting. The cell suspension was then transferred to tubes containing Wash medium [DMEM/F12 (Nacalai) supplemented with 7.5% BSA (Thermo)]. The cell suspension was centrifuged at 200 × g for 3 min at room temperature, and the supernatant was discarded. The cell pellet was resuspended in an appropriate volume of S10 medium supplemented with SCF, Y-27632, and Shield1. The cell suspension was added to the pre-coated culture plates in the desired amount, ensuring even distribution of cells (regarded as day 0). 3 × 104 cells were typically plated per well in a 24-well plate. The medium was replaced every 72 hours, with half of the medium exchanged for freshly prepared S10 medium containing SCF, Y-27632, and Shield. Starting from day 12, Shield1 was omitted from the fresh medium. SMART-seq stranded kit (TAKARA) was used.