Construction method of American paddlefish cardiac cell line
A construction method and technology of sturgeon, which can be applied to cell dissociation methods, artificial cell constructs, animal cells, etc., can solve problems such as diseases and economic losses in sturgeon breeding industry, and achieve a rapid proliferation rate, which is beneficial to cultivation and growth. Effect
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Embodiment 1
[0036] 1) Select a healthy sturgeon, first wash off the mucus and dirt on the fish's body surface with tap water, sterilize the tail with 75% ethanol, draw blood from the tail vein, filter it with a 0.22 μm microporous membrane, and take 250 μl of sturgeon serum, Add 15ml fetal bovine serum, add 3ml antibiotic mixed solution (penicillin mass concentration is 200IU / ml, streptomycin mass concentration is 200IU / ml, amphotericin B mass concentration is 0.5μg / ml), add DMEM basal culture solution to 50ml, which is to prepare the complete culture medium of the heart cells of the spoon kissed sturgeon. In order to promote the division and rapid proliferation of heart cells, 100 μl of human insulin-like growth factor type I, 100 μl of human epidermal growth factor, and 100 μl of human basic fibroblast-like growth factor were added to the above culture medium.
[0037] 2) The above-mentioned sturgeon was bathed in 20ppm potassium permanganate solution for 20 minutes, and then sterilized...
Embodiment 2
[0046] 1) Select a healthy sturgeon, first wash off the mucus and dirt on the fish's body surface with tap water, sterilize the tail with 75% ethanol, draw blood from the tail vein, filter it with a 0.22 μm microporous membrane, and take 250 μl of sturgeon serum, Add 15ml fetal bovine serum, add 3ml antibiotic mixed solution (penicillin mass concentration is 200IU / ml, streptomycin mass concentration is 200IU / ml, amphotericin B mass concentration is 0.5μg / ml), add DMEM basal culture solution to 50ml, which is to prepare the complete culture medium of the heart cells of the spoon kissed sturgeon. In order to promote the division and rapid proliferation of heart cells, 100 μl of human insulin-like growth factor type I, 100 μl of human epidermal growth factor, and 100 μl of human basic fibroblast-like growth factor were added to the above culture medium.
[0047] 2) The above-mentioned sturgeon was bathed in 20ppm potassium permanganate solution for 10 minutes, and then sterilized...
Embodiment 3
[0053] 1) Select a healthy sturgeon, first wash off the mucus and dirt on the fish's body surface with tap water, sterilize the tail with 75% ethanol, draw blood from the tail vein, filter and sterilize it with a 0.22 μm microporous membrane, and take 280 μl of sturgeon serum, Add 15ml fetal bovine serum, add 3ml antibiotic mixed solution (penicillin mass concentration is 200IU / ml, streptomycin mass concentration is 200IU / ml, amphotericin B mass concentration is 0.5μg / ml), add DMEM basal culture solution to 50ml, which is to prepare the complete culture medium of the heart cells of the spoon kissed sturgeon. In order to promote the division and rapid proliferation of heart cells, 100 μl of human insulin-like growth factor type I, 100 μl of human epidermal growth factor, and 100 μl of human basic fibroblast-like growth factor were added to the above culture medium.
[0054] 2) The above-mentioned sturgeon was bathed in 20ppm potassium permanganate solution for 15 minutes, and t...
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