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The rat ovary was chopped using scalpels into pieces of 0.3 mm3 and digested in DMEM/F12 (Thermo Fisher Scientific) containing 5% FBS,1 mg/mL collagenase IA (Sigma-Aldrich), 50 μg/mL Liberase and 1000 U DNase I (Roche, Sigma-Aldrich) in a shaking 37 °C incubator for 40 min 220rpm.Digestion was stopped with medium containing 10% FBS and cell suspension was centrifuged for 7 min on 300 × g.Discard the supernatant and cells were resuspend in DPBS, 2% FBS. The cell precipitate was passed through a 40 μm cell strainer (Millipore, USA).Red cell lysis solution(Solarbio, China) was added to the cell suspension for 5 min. Then the cell suspension was centrifuged for 5 min on 300 × g and resuspend in DPBS, 0.04%BSA.The cells were counted and the cell viability was calculated with Trypan Blue. |
RNA-Seq |
TRANSCRIPTOMIC SINGLE CELL |
PCR |
PAIRED
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