Serum-free culture solution for in-vitro culture of sheep ovarian cortex tissue
A serum-free culture medium and ovarian cortex technology, which is applied in the field of cell biology, can solve the problems that primordial follicles cannot be cultured in vitro for a long time, restrict industrial development, teaching and scientific research, etc., achieve clear system components, improve growth and development. The effect of survival time, simple preparation method
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[0030] 1. Configure culture medium
[0031] Basal culture medium: add α-MEM and DMEM at a volume ratio of 4:1.
[0032] Serum-free culture medium: add BSA 5 μg / mL, sodium pyruvate 0.2 mM, glutamine 3 mM, hypoxanthine 3 mM, insulin 10 μg / mL, transferrin 6.25 μg / mL to the above basal culture medium in sequence , sodium selenite 6.25 ng / mL, V C 50 μg / mL, penicillin 100 IU / mL, streptomycin 100 IU / mL.
[0033] 2. Obtain ovarian cortical tissue and fragments
[0034] The ovary tissue samples of Xinjiang military reclamation fine-haired lambs were collected by surgery, stored in 0.9% saline supplemented with 200 IU / mL penicillin and 200 IU / mL streptomycin at 37.5°C, and sent back to the laboratory within 1 h, and then added with Rinse twice in normal saline with 100 IU / mL penicillin and 100 IU / mL streptomycin, once in alcohol and twice in normal saline, and then remove the surface fascia and appendages of the ovary under a stereomicroscope.
[0035] Then under a stereomicroscope,...
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