Preparation method and application thereof for dog seminal fluid cryopreservation diluent
A cryopreservation and diluent technology, which is applied in applications, preservation of human or animal bodies, animal husbandry, etc., can solve the problems of low conception rate and litter size, improve survival rate, improve freeze resistance, and inhibit ice crystals the formation of the effect
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Embodiment 1
[0021] The quality ratio and preparation steps of the four diluent raw materials are as follows:
[0022] Diluent A: Vitamin E 1.2g, glucose 4g, citric acid 1.2g, fructose 2.5g, egg yolk 6g, penicillin 550,000 IU, streptomycin 0.13g, carnitine 0.2g, sodium glutamate 0.7g;
[0023] Diluent B: 3g glucose, 3ml DMSO, 12ml glycerol, 2ml ethylene glycol;
[0024] Diluent C: 0.8g trehalose, 1.5g gelatin, 1.2g EDTA, 0.5g sorbitol, 4g polyvinylpyrrolidone, 1.5g peptone;
[0025] Diluent D: Thimerosal 1.5g, Caspase inhibitor Z-VAD-FMK 4g.
[0026] The preparation methods of the four types of diluents are:
[0027] Diluent A: Put vitamin E, glucose, citric acid, fructose, egg yolk, carnitine, and sodium glutamate in a sterile beaker according to the weight ratio, dissolve in 100ml double distilled water, adjust the pH to 7.2, and stir 8min, then add penicillin and streptomycin according to the weight ratio, stir for 12min, store at 4℃;
[0028] Diluent B: Place glucose, DMSO, glycerol, and ethylene...
Embodiment 2
[0034] The quality ratio and preparation steps of the four diluent raw materials are as follows:
[0035] Diluent A: Vitamin E 1g, glucose 3g, citric acid 1g, fructose 2g, egg yolk 5g, penicillin 500,000 IU, streptomycin 0.1g, carnitine 0.1g, sodium glutamate 0.5g;
[0036] Diluent B: 2g glucose, 2ml DMSO, 10ml glycerol, 1ml ethylene glycol;
[0037] Diluent C: 0.5g trehalose, 1g gelatin, EDTA1, 0.3g sorbitol, 3g polyvinylpyrrolidone, 1g peptone;
[0038] Diluent D: 1.2 g of thimerosal and 3 g of caspase inhibitor Z-VAD-FMK.
[0039] The rest is the same as in Example 1.
Embodiment 3
[0041] The quality ratio and preparation steps of the four diluent raw materials are as follows:
[0042] Diluent A: Vitamin E 1.5g, glucose 5g, citric acid 1.5g, fructose 3g, egg yolk 7g, penicillin 600,000 IU, streptomycin 0.15g, carnitine 0.3g, sodium glutamate 1g;
[0043] Diluent B: 4g glucose, 4ml DMSO, 14ml glycerol, 3ml ethylene glycol;
[0044] Diluent C: 1.2g trehalose, 2g gelatin, 1.5g EDTA, 0.7g sorbitol, 5g polyvinylpyrrolidone, 2g peptone;
[0045] Diluent D: Thimerosal 1.8g, Caspase inhibitor Z-VAD-FMK 5g.
[0046] The rest is the same as Example 1
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