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Vitrification freezing method for sturgeon sperms

A technology of vitrification and sperm, which is applied in the preservation, application, and animal husbandry of human or animal bodies. It can solve the problems of high cost, impact on survival rate, and low application efficiency of biological cryopreservation technology, so as to improve the survival rate of cryopreservation. , good embryo development, and the effect of avoiding crystallization and recrystallization

Inactive Publication Date: 2014-07-23
HUNAN MIAOWANG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The existing semen cryoprotection method—slow freezing method is easy to form ice crystals, which will lead to the formation of extracellular ice and intracellular ice and damage the cells. In addition, the crystallization and recrystallization during the freezing and thawing process will affect its survival rate, making this biological cryoprotectant The storage technology is expensive and the application efficiency is extremely low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] A method for vitrification and freezing of sturgeon sperm, which is to uniformly dilute the sturgeon sperm to be frozen and the antifreeze protection solution in a ratio of 100:90 by volume, and then add 8 parts by volume of fresh egg yolk and 45 parts by volume Arachidonic acid and 6 parts by volume of glycerides are mixed, packed in freezer capsules, and frozen in a drawer type freezer at a freezing rate of -58 to -60°C / min. After freezing to -56 to -58°C, Then unload the capsule containing the semen mixture into a Dewar tank containing liquid nitrogen, control the freezing speed to be -148 to -152°C / min, and freeze it to -190 to -196°C before freezing.

[0016] The preparation method of the antifreeze protection solution is as follows: add 10% dimethyl phenol to the medical physiological saline according to the ratio and mix evenly, then add mannitol, and mix evenly; the weight ratio of the antifreeze protection solution is: Medical saline: 10% dimethylphenol: mannitol=...

Embodiment 2

[0018] A method for vitrification and freezing of sturgeon sperm, which is to uniformly dilute the sturgeon sperm to be frozen and the antifreeze protection solution in a ratio of 100:100 by volume, and then add 9 parts by volume of fresh egg yolk and 46 parts by volume Arachidonic acid and 7 parts by volume of glycerophospholipids are mixed into a semen mixture, which is packed in a freezer capsule and placed in a drawer-type freezer for freezing. The freezing rate is -60 to -62°C / min, and it is frozen to -60 to After -62℃, unload the capsule containing the semen mixture into a Dewar tank filled with liquid nitrogen, control the freezing rate to -148 to -152℃ / min, and freeze it to -190 to -196℃ and then freeze it .

[0019] The preparation method of the antifreeze protection solution is as follows: add 10% dimethyl phenol to the medical physiological saline according to the ratio and mix evenly, then add mannitol, and mix evenly; the weight ratio of the antifreeze protection sol...

Embodiment 3

[0021] A method for vitrification and freezing of sturgeon sperm, which is to evenly dilute the sturgeon sperm that needs to be frozen and the antifreeze protection solution in a ratio of 100:110 by volume, and then add 10 parts by volume of fresh egg yolk and 48 parts by volume. Arachidonic acid and 8 volume parts of lipoprotein are mixed into a semen mixture, which is packed in a freezer capsule and placed in a drawer-type freezer for freezing at a freezing rate of -64 to -66°C / min, and freezing to -64 to After -66℃, unload the capsule containing the semen mixture into a Dewar tank filled with liquid nitrogen, control the freezing rate to -148 to -152℃ / min, freeze it to -190 to -196℃ and store it .

[0022] The preparation method of the antifreeze protection solution is as follows: add 10% dimethyl phenol to the medical physiological saline according to the ratio and mix evenly, then add mannitol, and mix evenly; the weight ratio of the antifreeze protection solution is: Medic...

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PUM

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Abstract

The invention provides a vitrification freezing method for sturgeon sperms. The vitrification freezing method comprises the following steps: uniformly diluting the sturgeon sperms needing to be frozen and anti-freezing protection liquid according to a volume ratio of 100 to 90; adding following components in parts by volume: 8 parts of fresh egg yolk, 45 parts of arachidonic acid and 6 parts of glyceride; uniformly mixing to form seminal fluid mixed liquid; containing by a freezing capsule and putting the freezing capsule into a drawer type freezing room to be frozen at a freezing speed from -58 DEG C / min to -60 DEG C / min; after the freezing temperature reaches -56 DEG C--58 DEG C, unloading the capsule containing the seminal fluid mixed liquid into a Dewar flask; controlling the freezing speed ranging from -148 DEG C / min to -152 DEG C / min; and freezing to a temperature being -190 DEG C--196 DEG C, freezing and storing. According to the vitrification freezing method, the survival activity of the frozen sperms can be improved effectively; the unfrozen sperms are fertilized to fresh ova; the fresh ovum fertilization is close to the fresh sperm fertilization; the fertilization rate can reach 80-90% and embryos develop well.

Description

Technical field [0001] The invention relates to a deep-low temperature freezing and thawing technology for fish hybrid breeding, in particular to a method for vitrification and freezing of sturgeon sperm. Background technique [0002] The preservation of biological genetic diversity and the application of hybrid breeding, the selection in aquaculture-cryopreservation of sperm below 170 ℃ for fish reproduction, is the development trend of innovative applications of biotechnology. But at present, cryopreservation of fish sperm is only a method used in laboratories in highly developed countries, and the actual application range is very limited. The main reason is that many kinds of fish sperm will undergo significant damage during the freezing and thawing process. The degree of damage depends on the freezing rate, cryoprotectant and thawing method. [0003] Many scholars have proposed that if vitrification can be achieved during the cryopreservation of frozen semen, it will be more c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 赵智媛赵伟
Owner HUNAN MIAOWANG BIOTECH