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Refrigeration preservation and resuscitation method of sperm of non-human primates

A primate and cryopreservation technology, which is applied to the field of sperm cryopreservation and recovery of non-human primates, can solve problems such as poor recovery rate, and achieve a simple cryopreservation and recovery method, wide applicability, and price reduction. The effect of freezing costs

Active Publication Date: 2017-07-21
INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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AI Technical Summary

Problems solved by technology

Some researchers also used SpermCryo yolk-free freezing liquid to cryopreserve cynomolgus monkey sperm, and obtained 32.3% and 41.9% sperm cryopreservation recovery rates, but the recovery rate was still not good

Method used

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  • Refrigeration preservation and resuscitation method of sperm of non-human primates
  • Refrigeration preservation and resuscitation method of sperm of non-human primates

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Experimental program
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Effect test

Embodiment 1

[0019] Experimental animals: 6 healthy, fertile male cynomolgus monkeys and rhesus monkeys (3 each, aged 7-10 years) were from the Drug Safety Evaluation Research Center of the Institute of Medical Biology, Chinese Academy of Medical Sciences. Each monkey was raised in a single cage, fed with 3 meals a day, and given a certain amount of clean fresh fruit every day as supplementary food. Keep 12h / 12h light and dark every day, at room temperature of 17-22°C. The Drug Safety Evaluation Research Center of the Institute of Medical Biology, Chinese Academy of Medical Sciences has obtained the license for the use of experimental animals from the Kunming Science and Technology Bureau. The experimental protocol has been reviewed and approved by the Ethics Review Committee of the Institute of Medical Biology, Chinese Academy of Medical Sciences before implementation. The use of animals complies with the 3R (Reduction, Replacement, Refinement) principle. All operations related to exper...

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Abstract

The invention discloses a refrigeration preservation and resuscitation method of sperm of non-human primates. According to the refrigeration preservation and resuscitation method of sperm of non-human primates, in refrigeration preservation, sperm is subjected to liquefaction at normal temperature for 30min, and monkey sperm is diluted with HTF culture solution, wherein the volume ratio of the monkey sperm to the HTF culture solution is controlled to be 1:9; glycerin is added based on the volume of the monkey sperm until the volume final concentration of glycerin is 10%; an obtained mixture is subjected to subpackage in 1.5ml cryopreservation tubes, is stored at 4 DEG C for 30min, is stored for 30min in suspension above liquid nitrogen, and then is stored in liquid nitrogen; in resuscitation, the cryopreservation tubes stored in liquid nitrogen are shaken slightly in water bath at 37 DEG C for 1 to 3min until complete thawing is realized. Compared with the prior art, the survival rate and the fertility rate of the refrigeration preservation and resuscitation method are both increased; the refrigeration preservation and resuscitation method is effective, is rapid, and is convenient, and can be used for sperm refrigeration preservation and genetic resource conservation of other non-human primates.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a non-human primate sperm cryopreservation and recovery method. Background technique [0002] Non-human primates, especially cynomolgus monkeys and rhesus monkeys, are highly similar to humans in genetics, anatomy and physiology, making them widely used in the fields of life sciences and medical research. In terms of reproductive physiology, because these two animals have menstrual cycles similar to humans, this feature is often used to reveal human reproductive laws and seek ways and methods to control birth. With the establishment and widespread use of transgenic and gene knockout animal models, the establishment of their preservation and breeding systems is becoming more and more important. Among them, sperm freezing is in vitro fertilization (in vitro fertilization, IVF) or intracytoplasmic sperm microscopy. Injection (Intracytoplasmic Sperm Injection, ICSI) is an indispensable ...

Claims

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Application Information

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IPC IPC(8): A01N1/02C12N5/076
CPCA01N1/0221A01N1/0284C12N5/061
Inventor 马开利黄璋琼鲁帅尧李云江勤芳高家红
Owner INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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