Anti-senescence product for gonad granulocyte, its preparing method and uses
A granular cell, anti-aging technology, applied in biochemical equipment and methods, anti-toxic agents, microorganisms, etc., can solve the problems of not being able to simulate the physiological state well, the gap between physiological states is large, and the effective ingredients are single, so as to alleviate the symptoms of menopause, Convenient source, high bioavailability effect
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[0029] 1. Obtaining ovarian granulosa cells: 4-6 weeks old female Kunming mice were injected intraperitoneally with 7.5 U of pregnant horse serum (PMSG) per mouse, and they were killed by cervical dislocation after 48 hours of routine feeding. In a petri dish filled with 4°C pre-cooled balanced salt solution (BSS, containing 400IU / ml hyaluronidase, 100U / ml penicillin and streptomycin), puncture the follicles with a 28G injection needle under a stereomicroscope, The granular cells were released into the BSS; the pipette was blown repeatedly several times to disperse them into single cells and filtered through a 200-mesh filter, and the cells were washed twice by centrifugation.
[0030] 2. Acquisition of granulosa cell culture supernatant and cell extract: resuspend granulosa cells in DMEM / F12 medium containing 10% fetal bovine serum by volume, and adjust the cell density to 2×10 6 / ml inoculated culture flasks, cultured at 37°C and 5% CO2 by volume for 6 hours, then changed th...
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