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Method for bovine in-vitro fertilization embryo culture

A technology of embryo culture and embryo culture solution, which is applied in agriculture-animal husbandry and veterinary reproduction, cattle in vitro fertilization embryo culture, and related culture fluid in the application field of cattle in vitro fertilization embryo culture, which can solve the high cost and extensive restrictions of in vitro fertilization embryo production Application, low efficiency and other issues

Active Publication Date: 2018-11-06
天津力牧生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Although in vitro fertilization technology can be successfully applied to many mammals, the high cost and low efficiency of in vitro fertilized embryo production due to the low blastocyst rate of in vitro fertilization limit the wide application of this technology in the practice of rapid breeding of cattle

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Embodiment 1: the culture method of bovine in vitro fertilization embryo

[0055] 1. Reagents

[0056] In the specific test of the present invention, if not otherwise specified, the relevant reagents used are described in detail as follows:

[0057] Add double-antibody normal saline: normal saline containing 400 IU / mL penicillin and 400 μg / mL streptomycin.

[0058] Egg washing solution: BY basal culture solution supplemented with 3mg / mL bovine serum albumin.

[0059] Maturation medium: BY basal medium supplemented with 100 mL / L FBS, 10 μg / mL FSH, 10 μg / mL LH, 1 μg / mL E2, and 20 ng / mL EGF. Among them, EGF—epidermal growth factor, FSH—follicle stimulating hormone, FBS—fetal bovine serum, E2—estradiol, LH—luteinizing hormone.

[0060] Fertilization medium: containing 112.0mM sodium chloride, 4.02mM potassium chloride, 2.25mM calcium chloride dihydrate, 0.52mM magnesium chloride hexahydrate, 0.83mM potassium dihydrogen phosphate, 37.0mM sodium bicarbonate, 1.25mM pyru...

Embodiment 2

[0093] Embodiment 2: the culture method of bovine in vitro fertilization embryo

[0094] The main difference between this Example 2 and the above Example 1 is that an additional 0.25 mg / L sodium selenite and 0.075 mg / L anhydrous copper sulfate were added to the BY basal culture solution.

[0095] 1. Reagents

[0096] In the specific test of the present invention, if not otherwise specified, the relevant reagents used are described in detail as follows:

[0097] Add double-antibody normal saline: normal saline containing 400 IU / mL penicillin and 400 μg / mL streptomycin.

[0098] Egg washing solution: BY basal culture solution supplemented with 3mg / mL bovine serum albumin.

[0099] Maturation medium: BY basal medium supplemented with 100 mL / L FBS, 10 μg / mL FSH, 10 μg / mL LH, 1 μg / mL E2, and 20 ng / mL EGF. Among them, EGF—epidermal growth factor, FSH—follicle stimulating hormone, FBS—fetal bovine serum, E2—estradiol, LH—luteinizing hormone.

[0100] Fertilization medium: con...

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PUM

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Abstract

The invention relates to a method for bovine in-vitro fertilization embryo culture. The method comprises the following steps: collection of an oocyte and in-vitro maturity; in-vitro fertilization; embryo in-vitro culture and preservation. During the in-vitro fertilization, mature COCs are cultured in a fertilization culture solution of the invention, during embryo in-vitro culture, granule cells around an embryo are completely removed by using an egg peeling needle, the granule cells are placed into the embryo culture solution to be cultured, and after the culture is finished, the usable embryo is subjected to liquid nitrogen cryopreservation in refrigerating fluid. The method has many excellent technical effects as described in the description.

Description

technical field [0001] The invention belongs to the technical field of animal reproduction, and relates to a technology for agricultural-animal husbandry and veterinary reproduction, in particular to a method for culturing bovine in vitro fertilized embryos. In addition, the present invention also relates to the use of related culture fluids for the cultivation of bovine in vitro fertilized embryos. Application in in vitro fertilization embryo culture. Further, the present invention also relates to a method for culturing bovine IVF embryos using the relevant culture fluid for culturing bovine IVF embryos. In particular, the method for culturing bovine IVF embryos of the present invention has excellent technical effects. Background technique [0002] In Vitro Fertilization (In Vitro Fertilization) or (external fertilization) refers to the technology in which mammalian sperm and eggs complete the fertilization process in an artificially controlled environment outside the body...

Claims

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Application Information

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IPC IPC(8): C12N5/073C12N5/075C12N5/076A01N1/02
CPCA01N1/0221C12N5/0603C12N5/0609C12N5/061C12N2500/12C12N2500/14C12N2500/16C12N2500/32C12N2500/34C12N2500/35C12N2500/38C12N2500/40C12N2501/11C12N2501/30C12N2501/31C12N2501/392C12N2501/91C12N2501/999
Inventor 王小武郭晶王娜郝少强赵明礼马毅郭春明
Owner 天津力牧生物科技有限公司
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