Primary culture method of ampullaria gigas heart tissue cell
A tissue cell and cell culture technology, applied in the field of freshwater biological cell culture, can solve problems such as harm to crop production, outbreak of snails and other problems, and achieve the effect of strong repeatability and good culture effect.
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Embodiment 1
[0037] Preparation of cell culture medium
[0038] (Prepare 500mL):
[0039] Ingredient Quantity
[0040] RPMI1640 medium 400mL
[0041] Fetal bovine serum 50.0mL
[0042] Phenylthiourea saturated solution 50.0mL
[0043] Penicillin 100U
[0044] Streptomycin 0.1μg
[0045] Amphotericin B 0.25μg
[0046] In the aseptic operating table, mix the above ingredients and store at 4°C for later use.
Embodiment 2
[0047] Example 2 Primary culture of the heart tissue cells of the apple snail
[0048] Go through the following steps:
[0049] (1) Preparation of cell culture medium;
[0050] (2) Choose apple snails weighing 40 grams and raise them in sterile water for 12 hours;
[0051] (3) Scrub the mucus on the surface of the snail with a sterile cotton swab, and immerse the snail in 0.1% KMnO 4 solution, disinfect the surface for 10 minutes;
[0052] (4) Wash the apple snails 3 times with sterile distilled water;
[0053] (5) Blot the surface moisture of the snails with sterile filter paper;
[0054] (6) Place the snails in a dissection tray sterilized with 75% alcohol, and dissect the apple snails;
[0055] (7) Cut the heart tissue with dissecting scissors, put it into a six-well culture plate filled with HBSS cleaning solution and wash it 3 times, and cut it into 1mm with sterile scissors for the last time 3 small tissue blocks;
[0056] (8) Inoculate the treated tissue in step (7...
Embodiment 3
[0059] Example 3 Primary culture of the tissue cells of the apple snail core
[0060] Go through the following steps:
[0061] (1) Preparation of cell culture medium;
[0062] (2) Choose apple snails weighing 50 grams and raise them in sterile water for 12 hours;
[0063] (3) Scrub the mucus on the surface of the snail with a sterile cotton swab, and immerse the snail in 0.1% KMnO 4 solution, disinfect the surface for 15 minutes;
[0064] (4) Wash the apple snails 5 times with sterile distilled water;
[0065] (5) Blot the surface moisture of the snails with sterile filter paper;
[0066] (6) Place the snails in a dissection tray sterilized with 75% alcohol, and dissect the apple snails;
[0067] (7) Cut the heart tissue with dissecting scissors, put it into a six-well culture plate filled with HBSS cleaning solution and wash it 5 times, and cut it into 1mm with sterile scissors for the last time 3 small tissue blocks;
[0068] (8) Inoculate the treated tissue in step (...
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Abstract
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