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Beef cattle reproductive embryo cryopreservation liquid and freezing method

A cryopreservation solution and a freezing method technology are applied in the field of cryopreservation and freezing of beef cattle breeding embryos, which can solve the problems of poor cryopreservation effect and low freezing cell survival rate, and achieve improved cryopreservation effect and cell cryopreservation survival rate. , the effect of reducing the formation of ice crystals

Inactive Publication Date: 2019-12-24
内蒙古金草原牧业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention proposes a cryopreservation solution and freezing method for beef cattle reproductive embryos, which solves the problem in the prior art that the cryopreservation solution has a poor cryopreservation effect on cells, resulting in a low cryopreservation rate of cells

Method used

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  • Beef cattle reproductive embryo cryopreservation liquid and freezing method
  • Beef cattle reproductive embryo cryopreservation liquid and freezing method
  • Beef cattle reproductive embryo cryopreservation liquid and freezing method

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Bovine blastocyst freezing test

[0023] 1. Liquid preparation:

[0024] Pre-balanced solution: 45 parts of HPES buffer, 0.4 part of bovine serum albumin, 0.03 part of glutamine, 15 parts of polyethyleneimine, the preparation method of the pre-balanced solution is to stir and mix the components evenly at room temperature to obtain the pre-balanced solution. balance fluid.

[0025] Cryopreservation solution: 45 parts of HPES buffer, 0.3 parts of bovine serum albumin, 3 parts of sorbitol, 0.015 parts of glutamine, 12 parts of polyethyleneimine, 23 parts of sucrose, the preparation method of the cryopreservation solution is: in HPES First add polyethyleneimine to the buffer solution, add bovine serum albumin after dissolving, and let it dissolve at 4°C, then add sucrose, sorbitol and glutamine, stand at 4°C and dissolve to obtain a cryopreservation solution; before use , preheat the cryopreservation solution to 38°C.

[0026] Among them, the molecular weight of polyethy...

Embodiment 2

[0041] Bovine 2-cell embryo freezing test

[0042] 1. Liquid preparation:

[0043] Pre-balanced solution: 45 parts of HPES buffer, 0.4 part of bovine serum albumin, 0.03 part of glutamine, 15 parts of polyethyleneimine, the preparation method of the pre-balanced solution is to stir and mix the components evenly at room temperature to obtain the pre-balanced solution. balance fluid.

[0044] Cryopreservation solution: 40 parts of HPES buffer, 0.1 part of bovine serum albumin, 2 parts of sorbitol, 0.01 part of glutamine, 10 parts of polyethyleneimine, 20 parts of sucrose, the preparation method of the cryopreservation solution is: in HPES First add polyethyleneimine to the buffer solution, add bovine serum albumin after dissolving, and let it dissolve at 4°C, then add sucrose, sorbitol and glutamine, stand at 4°C and dissolve to obtain a cryopreservation solution; before use , first preheat the cryopreservation solution to 37°C.

[0045] Among them, the molecular weight of po...

Embodiment 3

[0061] Bovine 4-cell embryo freezing test

[0062] 1. Liquid preparation:

[0063] Pre-balanced solution: 45 parts of HPES buffer, 0.4 part of bovine serum albumin, 0.03 part of glutamine, 15 parts of polyethyleneimine, the preparation method of the pre-balanced solution is to stir and mix the components evenly at room temperature to obtain the pre-balanced solution. balance fluid.

[0064] Cryopreservation solution: 50 parts of HPES buffer, 0.5 part of bovine serum albumin, 5 parts of sorbitol, 0.02 part of glutamine, 15 parts of polyethyleneimine, 25 parts of sucrose, the preparation method of the cryopreservation solution is: in HPES First add polyethyleneimine to the buffer solution, add bovine serum albumin after dissolving, and let it dissolve at 4°C, then add sucrose, sorbitol and glutamine, stand at 4°C and dissolve to obtain a cryopreservation solution; before use , first preheat the cryopreservation solution to 40°C.

[0065] Among them, the molecular weight of po...

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Abstract

The invention belongs to the technical field of embryology. The invention provides beef cattle reproductive embryo cryopreservation liquid and a freezing method. The cryopreservation liquid is prepared from the following components in parts by weight: 40 to 50 parts of an HPES buffer solution, 0.1 to 0.5 part of bovine serum albumin, 2 to 5 parts of sorbitol, 0.01 to 0.02 part of glutamine, 10 to15 parts of polyethyleneimine and 20 to 25 parts of cane sugar. The beef cattle reproductive embryo freezing method comprises the following steps: balancing beef cattle embryos to be frozen in a pre-balancing solution for 5-8 minutes, balancing the beef cattle embryos in the beef cattle reproductive embryo freezing preservation solution according to any one of claims 1-3 for 30-40 seconds, transferring the beef cattle reproductive embryo freezing preservation solution to a freezing carrier, and putting the freezing carrier into liquid nitrogen for freezing treatment. By means of the technicalscheme, the problem that in the prior art, the cryopreservation effect of cryopreservation liquid on cells is poor, and consequently the cryopreservation survival rate of the cells is low is solved.

Description

technical field [0001] The invention belongs to the technical field of embryology, and relates to a cryopreservation solution and a freezing method for beef cattle breeding embryos. Background technique [0002] Embryo cryopreservation is to freeze the early embryos of animals with special protective agents and cooling measures, so that their metabolism stops or weakens to a small enough level in liquid nitrogen at -196°C without losing the ability to resume metabolism after warming up. ability to preserve embryos for a long period of time. Cryopreservation of bovine embryos was successfully achieved by Wilmut in 1973. According to different cooling methods, bovine embryo freezing methods can be divided into slow cooling method and vitrification freezing method. The slow cooling method refers to that the embryos treated with cryoprotectants are first cooled from room temperature at a rate of -1.0°C / min to -7°C at a rate of -7°C, and kept for 10 minutes. After freezing, co...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 张广志
Owner 内蒙古金草原牧业有限公司
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