Method for improving blastocyst rate of in-vitro embryos of animals

A technology of embryonic development and animal body, applied in embryonic cells, animal cells, vertebrate cells, etc., to reduce production costs, improve early development ability, and improve production efficiency.

Inactive Publication Date: 2012-10-03
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, how to improve the blastocyst develop

Method used

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  • Method for improving blastocyst rate of in-vitro embryos of animals
  • Method for improving blastocyst rate of in-vitro embryos of animals
  • Method for improving blastocyst rate of in-vitro embryos of animals

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 The effect of rbIFN-τ on the development of parthenogenetically activated embryos in bovine oocytes

[0023] 1. Collection and in vitro maturation of bovine oocytes

[0024] Bovine ovaries were collected from the slaughterhouse and placed in thermos flasks containing physiological saline at 27±2°C. After arriving at the laboratory, wash the ovaries with sterilized saline preheated at 37°C for 3 times, and then use a syringe to aspirate the cumulus-oocyte complexes (COCs) in the 2-8mm follicle above the ovary, and the complete cumulus cells will be collected. The layer or part of the dense cumulus cells was washed twice in the oocyte washing solution, and then washed three times in the maturation solution, and the washed oocytes were transferred into 100 μL oocyte maturation culture microdrops in vitro, and each microdrop was placed Insert 15~20 oocytes, and finally put them in 38.5℃, 5%CO 2 and cultured in a carbon dioxide incubator with saturated humidity f...

Embodiment 2

[0034] Example 2 The effect of rbIFN-τ on the development of bovine oocyte fertilized embryo in vitro

[0035] 1. In vitro fertilization of bovine oocytes

[0036] (1) Preparation of mature oocytes before in vitro fertilization

[0037] See item 1 in Example 1 for the collection and in vitro maturation of bovine oocytes. Transfer the oocytes matured in vitro for 24 hours into pre-balanced BO fertilization solution (38.5°C, 5% CO 2 (incubator) for 3 washes, and then transferred to 50 μL of fertilized fluid droplets in a pre-balanced 60 mm Petri dish and placed at 38.5 °C, 5% CO 2 15-20 mature oocytes per fertilized microdrop.

[0038] (2) Preparation of sperm

[0039] After thawing the frozen semen in a water bath at 37°C, take 0.25 μL of semen and carefully place it in a sterilized 10 mL centrifuge tube containing 7 mL of BO capacitation solution, then centrifuge (1800 rpm, 5 minutes) to wash twice, and then place under the microscope Adjust sperm concentration to 2×10 wi...

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Abstract

The invention provides a method for improving the blastocyst rate of in-vitro embryos of animals. According to the method, the developmental capacity of a parthenogenetic embryo and an in-vitro fertilization embryo can be remarkably improved by adding type I interferon into an embryonic development culture solution for the in-vitro culture of the embryos; compared with the blastocyst rate obtained under the culture of the conventional culture solution, the blastocyst rate is improved by 7 to 13 percent; and the blastocyst hatching rate is also remarkably higher than the blastocyst hatching rate under the culture of the conventional culture solution. By the method, the early development capacity of in-vitro embryos of cattle and sheep is improved, so that the in-vitro production efficiency of embryos of the cattle and sheep is improved, and high-quality embryos are provided for the scientific research and production field in which the embryos of the cattle and sheep are taken as materials.

Description

technical field [0001] The invention relates to the field of animal reproduction and breeding, in particular to a method for improving the blastocyst development rate of animal embryos in vitro. Background technique [0002] The in vitro production technology of embryos can not only improve the fecundity of animals, but also reduce production costs, and provide a large amount of materials for research on embryos. However, how to improve the blastocyst development rate is the difficulty of this technique. At present, domestic and foreign technologies to increase the development rate of bovine blastocysts in vitro mainly start from two aspects: improving the composition of the culture medium and the culture environment. For example, adding insulin, transferrin and sodium selenite (ITS) to oocyte maturation medium or adding leukemia growth inhibitory factor (LIF), basic fibroblast growth factor (bFGF) and Melatonin (MT) etc. can provide the blastocyst development rate of catt...

Claims

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Application Information

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IPC IPC(8): C12N5/073
Inventor 曾申明鲍忠剑赵栓
Owner CHINA AGRI UNIV
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