Method for adjusting and controlling ovarian granular cells based on circ-Atp5BP
A technology of granulosa cells and ovaries, applied in the field of ovarian granulosa cell regulation based on circ-Atp5BP, can solve the problems of tissue specificity and spatiotemporal specificity of circRNA, and achieve the effect of regulating proliferation.
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Embodiment 1
[0034] 1. Experimental plan
[0035] 1. Isolation and culture of porcine ovary granulosa cells
[0036] After the sows were slaughtered, the ovaries were taken out, washed in PBS, stored in saline at 38 degrees Celsius, and brought back to the laboratory within 1 hour to separate the ovarian granulosa cells.
[0037] Put the ovary in the preheated 37-degree normal saline and clean it, use a scalpel to cut the surface of the ovary evenly and lightly at intervals of 1mm, collect the follicle fluid, put the absorbed follicle fluid into a 15ml centrifuge tube, 1000rpm Centrifuge for 8 minutes, discard the supernatant, gently pipette with preheated PBS to evenly wash and centrifuge twice.
[0038] The prepared ovarian granulosa cell culture medium is 20% FBS plus DMEM and 1% third antibody (penicillin, streptomycin and amphotericin)
[0039] 2. Cell inoculation, subculture, transfection, and sample collection
[0040] When the ovarian granulosa cells proliferate to 85%, discard ...
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