Construction method of mandarin fish brain cell system
A construction method and technology of brain cells, applied in artificial cell constructs, methods based on microorganisms, animal cells, etc., can solve the problem of brain cell death due to hypoxia, no reports of brain-derived cell lines, and the difficulty of normal adhesion of primary brain cells wall proliferation
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[0040] 1) Treatment of brain tissue: First, soak and disinfect fresh mandarin fish with 75% alcohol for 2 minutes, wipe off the mucus on the body surface with gauze, dissect and remove the brain tissue in an ultra-clean workbench, and put it in a small chamber filled with HBSS balance solution. The brain tissue was roughly cut into tissue pieces of 50-100 mm3 in a glass bottle, collected by centrifugation at 600 rpm, and centrifuged repeatedly for 3 times.
[0041] 2) Primary culture: put the tissue block into the serum-containing collagenase digestion solution (digestion solution contains 10v / v% fetal bovine serum, 0.1w / v% type Ⅰ collagenase, 200U / ml penicillin, 200μg / ml ml streptomycin, 0.5 μg / ml amphotericin B in L-15 medium) for 6 hours at 28°C, after digestion, centrifuge at 1200 rpm for 5 minutes, collect the digested tissue cell suspension, Add 5 ml special proliferation culture medium for mandarin fish brain cells (the formula of culture medium is containing 20v / v% fet...
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