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A cryopreservation solution and cryopreservation technology, which is applied in the preservation, application, and animal husbandry of human or animal bodies, and achieves the effects of simple operation, good freezing effect, and easy promotion
Inactive Publication Date: 2018-04-06
GUANGDONG INST OF APPLIED BIOLOGICAL RESOURCES
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Semen cryopreservation methods and effectiveness vary widely among sturgeon species, study shows
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Embodiment 1～5
[0023] Formula per liter of Sturgeon's Cryopreservation Solution:
[0024] 20-30mM sucrose, 0.1-0.5mM potassium chloride, 20-40mM tris(hydroxymethyl)aminomethane hydrochloride (Tris-HCl), 5-10g / L bovine serum albumin (BSA), 2.5-10mM HCl Prototype glutathione (Glu), antifreeze ingredients (added by volume): 10% methanol (MT), or 15% dimethyl sulfoxide (DMSO), or 15% dimethyl sulfoxide (DMSO) + 10% fresh egg yolk, or 15% propylene glycol (PG), or 15% propylene glycol + 10% fresh egg yolk, and the rest is ultrapure water. Adjust the pH to 7.5-8.5 with HCl.
[0025] Preparation method:
[0026] Bovine serum albumin (BSA) 100g / L mother liquor: Weigh 10g of BSA powder, put it into a 200mL dry glass bottle, first add 80mL of ultrapure water, stir and dissolve on a magnetic stirrer, and then make up to volume in a volumetric flask to 100mL.
[0027] KCl 25mM mother liquor: Weigh 0.18g of KCl (molecular weight: 74.55) solid, put it into a 200mL dry glass bottle, first add 80mL of u...
experiment example
[0034] Taking liquid nitrogen ultra-low temperature (-196° C.) cryopreservation of 100 mL of sturgeon semen as an example, and taking Example 1 as an example, the application method of the sperm cryopreservation solution of Stryckeye's sperm is illustrated.
[0035] 1) Collection of fresh essence:
[0036]During the breeding season of Sturgeon's sturgeon, B-ultrasound examination is performed on the gonad development status of male broodstock, and large and mature male broodstock (139-158cm in total length) are selected for artificial induction, and luteinizing releasing hormone is injected intramuscularly at the base of the pectoral fin Analogue (LHRH), the injection dose is 2.5mg / kg, and the semen is collected after 36 hours. When collecting semen, use a dry towel to wipe around the genital opening of the broodstock, gently squeeze the abdomen of the broodstock, connect the semen to a 500mL sterile airtight plastic bottle through a catheter with a diameter of 1-2cm, and imme...
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Abstract
The invention discloses an acipenser schrenckii cryopreservation fluid and a long-term acipenser schrenckii sperm cryopreservation method. The pH of each liter of acipenser schrenckii cryopreservationfluid is 7.5-8.5. The acipenser schrenckii cryopreservation fluid is prepared from 20-30 mM of sucrose, 0.1-0.5 mM of potassium chloride, 20-40 mM of Tris-HCl, 5-10 g/L of bovine serum albumin, 2.5-10mM of reduced glutathione and antifreeze components and the balance ultra-pure water, wherein the antifreeze components are, by volume, one of 10% of methanol, 15% of DMSO, the mixture of 15% of DMSOand 10% of fresh egg yolk, 15% of propylene glycol and the mixture of 15% of propylene glycol and 10% fresh egg yolk mixture. The components of the acipenser schrenckii cryopreservation fluid are clear and definite, can protect sperms against damage caused by ice crystal, oxidization and the like, the fertilization rate can reach 45.87% after cryopreserved sperms are unfrozen and revive, healthyfry can be hatched, and the acipenser schrenckii cryopreservation fluid has the advantages of being simple, convenient, easy to operate and capable of completing cryopreservation without precious instruments when being applied to long-term acipenser schrenckii sperm cryopreservation.
Description
technical field [0001] The invention belongs to the technical field of low-temperature cryopreservation of biology, and more specifically, the invention relates to a cryopreservation liquid of Strykyu sturgeon sperm, a preparation method thereof and an application in a long-term cryopreservation method of Strykyu sturgeon sperm. Background technique [0002] Sturgeons originated in the Devonian period (450-350 million years ago), and are the only existing order in the cartilaginous sclerophyta, known as "living fossils". At present, there are 27 species of sturgeon, which are divided into two families, one is the family Acipenseridae, which includes 25 species, and the other is the family Acipenseridae, which includes 2 species. Sturgeon roe sauce is one of the world's most famous "luxury" foods of animal origin, and is known as "black gold". Therefore, human factors such as overfishing have caused a significant decline in the number of wild sturgeon. At present, in the lat...
Claims
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Application Information
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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 张秀娟陈金平吴文化马荆鄂李林妙江海英
Owner GUANGDONG INST OF APPLIED BIOLOGICAL RESOURCES
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