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Cryoprotectant and ultralow-temperature cryopreservation method for stichopus japonicus sperms

A technology of cryoprotectant solution and sperm, which is applied to the preservation, application, and animal husbandry of human or animal bodies. It can solve the problems of japonicus germplasm degradation and asynchronous maturation of male and female sea cucumbers in the north and south, so as to achieve a large amount of cryopreservation and solve unresolved problems. Simultaneous maturity, objective and credible data results

Active Publication Date: 2020-12-15
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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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 ultralow-temperature cryopreservation of sea cucumber germplasm resources, in particular to a cryoprotectant, an activator formula and an ultralow-temperature cryopreservation for apostichopus japonicus sperm method based on a program cooling instrument and CASA. The cryoprotectant is composed of a diluent, an antifreeze agent and an additive. The diluent is natural seawater, the antifreeze agent is one or more of aprotic polar solvents, and the additive is glucose. According to the method, the freezing stock is large (more than 200 mL of fresh stichopus japonicus seminal fluid can be frozen in one batch), the cooling process can be precisely controlled, the application effect is good after thawing, and the problems of asynchronous maturationof male and female sea cucumbers in south and north and a series of stichopus japonicus germplasm degradation in actual production can be solved.

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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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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