Low temperature preservation method of sperm pods of Litopenaeus vannamei
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A technology of cryopreservation and preservation method, which is applied to the preservation, application, and animal husbandry of human or animal bodies. It can solve the problems of difficult sperm fertilization, impact on sperm vitality, and a survival rate of only 60%, achieving a high sperm survival rate. , simple operation, save the complete effect
Inactive Publication Date: 2011-11-30
GUANGXI INST OF FISHERIES
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Although this method preserves the sperm, it has a certain impact on the motility of the sperm during the process of processing the sperm pods, and it is difficult to fertilize the sperm after the sperm pods have been removed, and the survival rate is only 60%.
Therefore, this method cannot fully meet the conditions for subsequent research work using preserved sperm
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Embodiment 1
[0019] One, fixing and providing the preparation of the nutrient substance culture layer required for sperm pods:
[0020] (1) Culture medium:
[0021] Dissolve 1 bag of 1640 culture medium from American GIBCO Company in 500ml of seawater with a salinity of 30‰, which was sterilized by filtration, to prepare 2×1640 culture medium with a concentration of 20.8g / L, and add 2.38g of HEPES (hydroxyl Ethylpiperazine (ethanesulfonic acid) buffering agent in the solution to its final concentration is 10mmol / L, adding aseptic 1mol / L NaOH to adjust pH 7.2, adding penicillin and streptomycin to adjust its final concentration is 1 million units, Then filter through a 0.22μm filter and pack into 50mL / bottle for later use.
[0022] (2) Self-made type I liquid rat tail collagen:
[0023] Take 1 rat tail, soak it in 75% alcohol for 30 minutes, tear off the rat tail skin under aseptic conditions, cut the tail into several sections, pull out the white tail tendon, and immerse it in 150ml of 0...
Embodiment 2
[0029] One, fixing and providing the preparation of the nutrient substance culture layer required for sperm pods:
[0030] (1) Culture medium:
[0031] Dissolve 1 bag of 1640 culture medium from American GIBCO Company in 500ml of seawater with a salinity of 30‰, which was sterilized by filtration, to prepare 2×1640 culture medium with a concentration of 20.8g / L, and add 2.38g of HEPES (hydroxyl Ethylpiperazine (ethanesulfonic acid) buffer in the solution to a final concentration of 10mmol / L, adding sterile 1mol / L NaOH to adjust the pH to 7.3, adding penicillin and streptomycin to adjust the final concentration to 1 million units , and then filtered through a 0.22μm filter and divided into 50mL / bottles for later use.
[0032] (2) Self-made type I liquid rat tail collagen:
[0033] Take 1 rat tail, soak it in 75% alcohol for 30 minutes, tear off the rat tail skin under aseptic conditions, cut the tail into several sections, pull out the white tail tendon, and immerse it in 150...
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Abstract
The invention discloses a method for storing spermatophore of litopenaeus vannamei at low temperature. The method comprises the following steps of: acquiring the spermatophore, fixing the spermatophore, storing the spermatophore and dyeing spermatozoa. In the method, a culture layer for the spermatophore of the litopenaeus vannamei is prepared from 0.3 to 0.4 part of 1.8 to 3.0 percent self-prepared type I liquid rat tail collagen serving as a medium for contracting the culture layer, 0.3 to 0.4 part of culture solution of 2*1,640, 0.1 to 0.2 part of fetal calf serum in an American GIBCO company and 0.1 part of Matrigel (a soluble basilar membrane matrix) in an American Sigma company, wherein the culture layer is viscous at normal temperature, and can be solidified at the temperature of 40 DEG C and can wrap the spermatophore. The spermatophore is packaged in separate an epoxy resin (EP) tube at the normal temperature, the spermatophore is wrapped by the prepared culture layer, and the EP tube wrapped with the spermatophore is arranged in a refrigerator to be stored at the low temperature of 4 DEG C. The method has the characteristics of simplicity of operation, complete storage of a spermatophore structure, high survival rate of the spermatozoa, long retention time, stable effect of low-temperature storage and the like and is easy to popularize and apply. The retention time of the spermatophore is at least 3 months, and the survival rate of the spermatozoa is over 80 percent.
Description
technical field [0001] The invention relates to a method for low-temperature preservation of animal sperm, in particular to a method for low-temperature preservation of sperm pods of Litopenaeus vannamei. Background technique [0002] Litopenaeus vannamei, also known as Penaeus vannamei, is one of the large-scale cultured prawn species in the world and is the most productive prawn in my country. Although more and more important in the breeding industry, its sperm preservation technology is still rare, which greatly limits the feasibility of selective breeding. At present, the method of cryopreservation of sperm of mammals and fish mostly adopts ultra-low temperature freezing method of liquid nitrogen, and the cryopreservation technology of sperm has the disadvantages of low survival rate of frozen sperm, low fertilization rate, and small storage capacity of frozen sperm, which is not practical. level and many other issues, and because the sperm of the prawn is different fro...
Claims
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Application Information
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