Porcine ovarian granulosa cell primary culture method
A technology of granulosa cells and ovaries, applied in cell dissociation methods, animal cells, tissue culture, etc., can solve the problems of impure cell survival rate and time-consuming, and achieve the goal of shortening in vitro operation time, improving survival rate and avoiding pollution Effect
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[0026] The present invention will be specifically introduced below in conjunction with the accompanying drawings and specific embodiments.
[0027] 1. Prepare the solution
[0028] Solution I: 1×PBS solution.
[0029] Solution II: DMEM / F12 culture solution containing 1×antibiotics.
[0030] Solution III: Cell complete growth medium.
[0031] The preparation method of the complete cell growth culture medium (Solution III): mix the DMEM / F12 culture medium and FBS according to the volume ratio of 9:1, then add 1× antibiotics, and mix evenly.
[0032] 2. Collection of ovaries
[0033] Collect ovaries from puberty gilts at a local slaughterhouse, place the ovaries in an insulated container filled with sterile solution I at 37°C, and send them to the laboratory within two hours.
[0034] 3. Extraction of follicular fluid
[0035] Use a 20ml, 18g needle syringe to insert a needle into the follicle at a distance of 0.5mm from the base of the follicle, aspirate the follicle fluid,...
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