Red sea bream embryo freezing preservation method

A cryopreservation, embryo technology, applied in the preservation, application, animal husbandry, etc. of human or animal body

Inactive Publication Date: 2005-03-16
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

The cryopreservation methods for fish embryos mentioned above are not applicable to the preservation of sea bream embryos

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0016] 1) Take the fertilized eggs of the same batch of naturally produced red sea bream, place them in a slightly inflated hatching bucket with a seawater temperature of 18±1°C, and cultivate them until the embryos are at the heartbeat stage, and the heartbeat frequency is more than 60 times / min;

[0017] 2) Fish out the embryos with 80-mesh sieve silk net, lightly soak the surface seawater with filter paper, put it into DMSO5%+PG5% antifreeze solution with balanced salt solution Cortland as the base liquid, and soak in the refrigerator at 0-4 °C 90 minutes (pre-cooling); take two 2ml cryopreservation tubes from the refrigerator, each absorb 1.2ml of antifreeze, one of which contains 61 embryos, and the other contains 62 embryos; quickly put it into the cooling device, start to cool down;

[0018] 3) The cooling program is: 0~-15.5°C, the cooling rate is 3.5°C / min; -15.5~-30°C, the cooling rate is 3°C / min; when the temperature is -12°C, quickly take it out of the cooling devi...

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PUM

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Abstract

The invention relates to a refrigeration method for preserving sea fish embryo which comprises, selecting heartbeat stage embryos, charging into 0-4 deg C. antifreezing liquid, pre-cooling for 90-60 minutes, sectionwise cooling down, placing cores at -12 deg C during the cooling period, preserving in liquid nitrogen at the temperature of -30 deg C, equalizing 10 minutes at -80 deg C when defreezing, defreezing with 43 deg C water till full thawing, eluting with eluent, cultivating in sea water so as to obtain the revival embryos.

Description

technical field [0001] The invention relates to cryopreservation of fish embryos, in particular to a method for cryopreservation of marine fish red sea bream embryos. The red sea bream embryos are cooled, stored in liquid nitrogen, thawed and eluted to obtain revived embryos. Background technique [0002] The cryopreservation of fish embryos has a history of about 20 years. Due to the large size of fish embryos, high water content, high yolk content, and poor egg membrane permeability, cryopreservation is much more difficult than mammalian embryos. Scholars at home and abroad have done a lot of research on the damage mechanism of fish embryo cryopreservation, freezing and cooling rate, thawing and rewarming, antifreeze types and toxicity, and have made great progress. At present, the cryopreservation of Cyprinidae embryos at temperatures above -100°C has achieved a relatively stable resurrection rate [Zhang Kejian, Lou Yundong, Zhang Yinjiang et al. Cryopreservation of thre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 李军肖志忠丁福红马道远徐世宏
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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