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Ultra low temperature refrigeration method for preserving sperm of sturgeon

An ultra-low temperature freezing, fish sperm technology, applied in the preservation, application, animal husbandry and other directions of human or animal body, can solve the problems of low survival rate of frozen sperm, unstable preservation effect, and no sperm osmotic pressure value measured. , to avoid errors

Inactive Publication Date: 2005-03-30
EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

[0002] Acipenseridae are rare and precious aquatic species. The preservation of sturgeon spermatozoa is related to the protection of rare species. However, there is still no successful method for preserving the spermatozoa of Acipenseridae.
[0003] In the prior art, liquid nitrogen containers are used to store the sperm of silver carp, bighead carp, grass carp, and carp freshwater fish. The temperature of liquid nitrogen is -196°C. The preservation effect is unstable. At the same time, the optimal osmotic pressure value for sperm cryopreservation has not been measured in cryopreservation technology, and the survival rate of frozen sperm is not high

Method used

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Embodiment Construction

[0007] The present invention is equipped with an FM-8P freezing point osmometer to measure the osmotic pressure of the sperm of Acipenseridae, for example, the osmotic pressure of the sperm of Acipenseridae is measured to be 54-56mOsm / kg, and the sperm preservation solution is prepared according to the osmotic pressure value , the sperm preservation solution is prepared with NaCl, KCl, and glucose, and the concentration of NaCl can be 0.3%, the concentration of KCl is 0.03%, and the concentration of glucose is 1.2%. The concentration of glucose is 1.2%, or 100ml of solution contains NaCl0.5g, KCl0. 03g, glucose 1.2g preservation solution; DMSO was used as antifreeze agent, the concentration was 12%; the ratio of sperm and protection solution was diluted 3:1; after 2-3 hours of equilibrium treatment at a low temperature of 2-4°C, the sperm Put it into a 2ml plastic centrifuge tube, and cool it through a low-temperature biological cooling device; lower it to -5°C for 10 minutes, ...

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Abstract

A method for ultralow-temp freeze storage of sturgeon sperm with high activity includes measuring the osmotic pressure of sturgeon sperm, determining the antifreeze protective liquid, preparing it by mixing liquid diluent with DMSO, proportionally mixing sperm with said protective liquid, low-temp balancing, loading in centrifugal tube, lowering temp to -5 deg.C, balancing, putting in liquefied nitrogen at -160- -180 deg.C, balancing, storing it in liquefied nitrogen, and two-step thawing: raising temp for -196 deg.C to -160- -180 deg.C and balancing and thawing by water bath at 38-40 deg.C.

Description

Technical field: [0001] The invention relates to a method for cryopreserving fish sperm. Background technique [0002] Acipenseridae is a rare and precious aquatic species, and the preservation of spermatozoa in Acipenseridae is related to the protection of rare species. However, there is still a lack of successful methods for preserving spermatozoa in Acipenseridae. [0003] In the prior art, liquid nitrogen containers are used to store the sperm of silver carp, bighead carp, grass carp, and carp freshwater fish. The temperature of liquid nitrogen is -196°C. The preservation effect is unstable. At the same time, the optimal osmotic pressure value for sperm cryopreservation has not been measured in cryopreservation technology, and the survival rate of frozen sperm is not high. Contents of the invention [0004] The technical problem to be solved by the present invention is to provide a cryopreservation method that can preserve the sperm of sturgeons and has a relatively h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/00
Inventor 章龙珍庄平张涛黄晓荣冯广朋
Owner EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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