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A cryoprotectant solution and ultra-low temperature cryopreservation method for japonicus japonicus sperm

A cryoprotectant liquid and sperm technology, which is applied in the preservation, application, and animal husbandry of human or animal bodies, can solve the problems of asynchronous maturation of male and female sea cucumbers in the north and south, and degradation of japonicus germplasm, so as to achieve a large amount of frozen storage and production Convenience, very low cost effect

Active Publication Date: 2021-08-31
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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Problems solved by technology

[0004] The purpose of the present invention is to solve the above-mentioned problems existing in the prior art and solve the problems of asynchronous maturation of male and female sea cucumbers in the north and south and a series of germplasm degradation problems of sea cucumbers in actual production. Liquid formula, activator formula and cryopreservation method of japonicus japonicus sperm

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] a. Prepare cryopreservation solution: prepare fresh ultra-low temperature cryoprotectant solution according to the principle of ready-to-use. ≥99.97% dimethylsulfoxide (DMSO), 1.98g glucose; formula (2) 80mL natural seawater filtered with a 0.45μm filter membrane, 20mL dimethylacetamide (DMA) with a purity≥99.97%. Pre-cool the prepared cryoprotectant solution in a constant temperature refrigerator at 4°C until use.

[0024]b. Semen acquisition: take sea cucumbers in the reproductive period, and dry the body after they discharge a large amount of water in the body cavity (avoid contact with seawater to activate the sperm). Use a sterile scalpel to dissect the sea cucumber from the abdomen to avoid scratching the gonads, pick out the complete gonads with tweezers, blot the body fluid on the surface of the gonads with absorbent paper, transfer them to a sterile petri dish, cut them into pieces with scissors, and clean them with a 300 mesh Sieve it into a sterile 50ml cent...

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Abstract

The invention relates to the technical field of ultra-low temperature cryopreservation of sea cucumber germplasm resources, in particular to a cryoprotectant solution, an activator formula, and a method for ultra-low temperature cryopreservation of imitation japonicus japonicus sperm based on a programmed cooling instrument and CASA. The cryoprotective solution is composed of diluent, antifreeze and additives; wherein, the diluent is natural seawater, the antifreeze is one or more of aprotic polar solvents, and the additive is glucose. This method has a large amount of frozen storage (more than 200mL of fresh japonicus semen can be frozen in one batch), can precisely control the cooling process, and has a good application effect after thawing. quality degradation problem.

Description

technical field [0001] The invention relates to the technical field of ultra-low temperature cryopreservation of sea cucumber germplasm resources, in particular to a cryoprotectant solution, an activator formula, and a method for ultra-low temperature cryopreservation of imitation japonicus japonicus sperm based on a programmed cooling instrument and CASA. Background technique [0002] Apostichopus japonicus, also known as sea cucumber, belongs to the phylum Echinodermata, class Holothuroidea, order Aspidochirotida, family Stichopodidae, genus Apostichopus, natural habitat It is the most common marine higher invertebrate in the northern part of the Western Pacific Ocean, including the coasts of the Far East of Russia, the coasts of Japan and South Korea, and the Yellow and Bohai Seas of China. Sea cucumber is a nourishing product with the same origin of medicine and food. Among the more than 20 kinds of edible sea cucumbers distributed in my country's sea area, the nutrition...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226A01N1/0284A01N1/021
Inventor 张立斌许帅刘石林林承刚孙景春孙丽娜杨红生
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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