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Bovine oocyte in-vitro fertilization embryo culture method and used culture medium

A technology of oocytes and culture medium, applied in the direction of cell culture active agents, embryonic cells, animal cells, etc., can solve the problems of high production cost, low efficiency, and limited wide application of in vitro fertilized embryos

Active Publication Date: 2018-11-02
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

[0062] Embodiment 1: the culture method of bovine in vitro fertilization embryo (no selenium in vitro)

[0063] 1. Reagents

[0064] In the specific test of the present invention, if not otherwise specified, the relevant reagents used are described in detail as follows:

[0065] Add double-antibody normal saline: normal saline containing 400 IU / mL penicillin and 400 μg / mL streptomycin.

[0066] Egg washing solution: BY basal culture solution supplemented with 3mg / mL bovine serum albumin.

[0067] 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.

[0068] HEPES maturation medium: BY basal medium supplemented with 15mmol / L HEPES, 100mL / L FBS, 10μg / mL FSH, 10μg / mL LLH, 1μg / mL E2, and 20ng / mL EGF.

[0069] Among them, EGF—epidermal growth factor, FSH—follicle stimulating hormone, FBS—fetal bovine serum, E2—estradiol, LH—luteinizing hormone.

[0070] Fertilization medium: containing 112.0mM sodium chlori...

Embodiment 2

[0103] Embodiment 2: the culture method of bovine in vitro fertilization embryo (living body without selenium)

[0104] 1. Reagents

[0105] In the specific test of the present invention, if not otherwise specified, the relevant reagents used are described in detail as follows:

[0106] Add double-antibody normal saline: normal saline containing 400 IU / mL penicillin and 400 μg / mL streptomycin.

[0107] Egg washing solution: BY basal culture solution supplemented with 3mg / mL bovine serum albumin.

[0108] 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.

[0109]HEPES maturation medium: BY basal medium supplemented with 15mmol / L HEPES, 100mL / L FBS, 10μg / mL FSH, 10μg / mL LLH, 1μg / mL E2, and 20ng / mL EGF.

[0110] Among them, EGF—epidermal growth factor, FSH—follicle stimulating hormone, FBS—fetal bovine serum, E2—estradiol, LH—luteinizing hormone.

[0111] Fertilization medium: containing 112.0mM sodiu...

Embodiment 3

[0144] Embodiment 3: the culture method of bovine in vitro fertilization embryo (isolated selenium copper)

[0145] The main difference between this example and the above example 1 is that 0.25 mg / L sodium selenite and 0.075 mg / L anhydrous copper sulfate are additionally added to the BY basal culture solution.

[0146] 1. Reagents

[0147] In the specific test of the present invention, if not otherwise specified, the relevant reagents used are described in detail as follows:

[0148] Add double-antibody normal saline: normal saline containing 400 IU / mL penicillin and 400 μg / mL streptomycin.

[0149] Egg washing solution: BY basal culture solution supplemented with 3mg / mL bovine serum albumin.

[0150] 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.

[0151] HEPES maturation medium: BY basal medium supplemented with 15mmol / L HEPES, 100mL / L FBS, 10μg / mL FSH, 10μg / mL LLH, 1μg / mL E2, and 20ng / mL EGF. ...

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Abstract

The invention relates to a bovine oocyte in-vitro fertilization embryo culture method and a used culture medium. The basic culture medium is an aqueous solution comprising sodium chloride, sodium dihydrogen phosphate, sodium bicarbonate, sodium acetate and the like. The invention further relates to an ovum washing solution and a mature culture solution. The bovine oocyte in-vitro fertilization embryo culture method comprises the following steps: collection and in-vitro maturation of an oocyte; in-vitro fertilization; and in-vitro culture and storage of the embryo. The bovine oocyte in-vitro fertilization embryo culture method has the excellent technical effect as shown in specification.

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. In particular, the present invention relates to a method for in vitro fertilization and embryo culture of live or isolated bovine oocytes. Background technique [0002] In Vitro Fertilization (In Vitro Fertilization) or (external fertilization) refers to th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073A01N1/02
CPCA01N1/0226C12N5/0604C12N2500/12C12N2500/14C12N2500/16C12N2500/32C12N2500/34C12N2500/38C12N2500/40C12N2501/11C12N2501/31
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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