Functionalized kit for preparing hydrogels
A kit and activator technology, applied in the field of sperm sustained-release composition, can solve problems such as lack of a large amount of research, means of preparation steps of centralized preservation system, etc.
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Embodiment 1
[0194] Example 1: Preparation of bovine spermatozoa in an alginate gel in an insemination straw
[0195] Material
[0196] The following chemicals were used: Tris hydrochloride, EDTA, NaHCO from Sigma-Aldrich (St. Luis, USA) 3 , NaCl, NaOH, glycerol (>99%) fructose, ascorbic acid and sodium citrate, carboxymethylcellulose (CMC), polyvinyl alcohol (PVA) from Sekisui Specialty Chemicals Europe S.L (Tarragona Spain), Selvol 523 / Selvol 325 . Anhydrous dextrose from Apro (Oslo, Norway). ViCality AlbaFil calcium carbonate from Brenntag Specialties (South Plainfield, USA) and sodium alginate (UP-LVG) from Novamatrix A / S (Drammen, Norway).
[0197] source of sperm
[0198] Bovine sperm in Trondheim, Norway and Stange's Store Ree's Geno device.
[0199] buffer
[0200] The following supplement solutions were used:
[0201] Supplement for the first dilution of sperm: 1.45g l -1 Tris hydrochloride glucose, 0.4g l -1 Sodium citrate, 1g l -1 Fructose, and 200ml l -1 egg yo...
Embodiment 2
[0212] Example 2: Determination of the mechanical strength of different alginate gels with immobilized sperm cells
[0213] Material
[0214] The following chemicals were used: Tris hydrochloride, EDTA, D-(+)-glucono delta-lactone, NaHCO from Sigma-Aldrich (St. Louis, USA) 3 , NaCl, glycerin (>99%), fructose and sodium citrate. Anhydrous dextrose from Apro (Oslo, Norway). Eskal 500 calcium carbonate from KSL staubtechnik GmbH (Lauingen, Germany) and sodium alginate (UPLVG and UP VLVG) from NovaMatrix, FMC BioPolymer AS (Sandvika, Norway). 0.25 ml standard insemination pipette (French micropipettor (IMV, L'Aigle, France)).
[0215] source of sperm
[0216] Bovine sperm in Trondheim, Norway and Stange's Store Ree's Geno device.
[0217] buffer
[0218] The following supplement solutions were used:
[0219] Supplement for the first dilution of sperm: 1.45g l -1 Tris hydrochloride dextrose, 0.4gl -1 Sodium citrate, 1g l -1 Fructose, and 200ml l -1 egg yolk. The...
Embodiment 3
[0225] Example 3: Coated insemination straws
[0226] Preparation of coated insemination straws
[0227] Materials: Polyvinyl alcohol Selvol 523 and Selvol 325 from Sekesui Specialty Chemicals Europe S.L. (Tarragonia, Spain), Carboxymethylcellulose (CMC) from CP Kelco: CEKOL, cellulose gum, sodium carboxymethylcellulose E466, A mixture of methylcellulose and hydroxypropylmethylcellulose Benecel MP 812W from Ashland Wilmington (Delaware USA), hypromellose Methocel K100 from Dow Chemical Company, ascorbic acid from Sigma Aldrich, AnalaR hydrochloride from VWR
[0228] 1) Coat the insemination straw by rinsing with a mixture of 13.6 μmol / mL aqueous ascorbic acid and 3% PVA (polyvinyl alcohol) aqueous solution or 5% PVA (polyvinyl alcohol).
[0229] 2) The insemination straw was coated by rinsing with a mixture (coating solution) of a 13.6 μmol / mL aqueous ascorbic acid solution and a 1.5% CMC (carboxymethylcellulose) aqueous solution. After rinsing with the coating solution,...
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