Method for promoting cow in-vitro embryo culture germ cell development

A technology of embryo culture medium and culture medium, which is applied to promote the in vitro maturation of oocytes, agricultural-animal husbandry and veterinary reproduction, in vitro fertilization and embryo culture, and the application of related culture medium in the cultivation of bovine in vitro fertilized embryos to promote the development of fertilized eggs, In the field of bovine in vitro fertilized embryo culture, it can solve the problems of low efficiency, high production cost of in vitro fertilized embryos, and restrictions on wide application

Inactive Publication Date: 2019-07-30
天津力牧鼎丰生物技术有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Although in vitro fertilization technology can be successfully applied to many mammals, the high cost and low efficiency of in vitro fertilized embryo

Method used

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  • Method for promoting cow in-vitro embryo culture germ cell development

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1: the culture method of bovine in vitro fertilization embryo (no selenium in vitro)

[0054] 1. Reagents

[0055] In the specific test of the present invention, if not otherwise specified, the relevant reagents used are described in detail as follows:

[0056] Add double-antibody normal saline: normal saline containing 400 IU / mL penicillin and 400 μg / mL streptomycin.

[0057] Egg washing solution: BY basal culture solution supplemented with 3mg / mL bovine serum albumin.

[0058] 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.

[0059] 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.

[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...

Embodiment 2

[0093] Embodiment 2: the culture method of bovine in vitro fertilization embryo (living body without selenium)

[0094] 1. Reagents

[0095] In the specific test of the present invention, if not otherwise specified, the relevant reagents used are described in detail as follows:

[0096] Add double anti-physiological saline, egg washing solution: the same as in Example 1.

[0097] 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.

[0098] 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.

[0099] Fertilization culture fluid, semen preparation culture fluid, embryo culture fluid, BY basal culture fluid: the same as in Example 1.

[0100] 2. Cattle in vitro fertilization and embryo culture:

[0101] Step (1), collection and in vitro maturation of oocytes

[0102] Live bovine eggs were collected, and the obtain...

Embodiment 3

[0129] Embodiment 3: the culture method of bovine in vitro fertilization embryo (isolated selenium copper)

[0130] 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.

[0131] 1. Reagents

[0132] In the specific test of the present invention, if not otherwise specified, the relevant reagents used are described in detail as follows:

[0133] Add double anti-physiological saline, egg washing solution: the same as in Example 1.

[0134] 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.

[0135] 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.

[0136] Fertilization culture fluid, semen preparation culture fluid, embryo culture fluid: the same as in Example 1.

[0137...

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Abstract

The invention relates to a method for promoting cow in-vitro embryo culture germ cell development. On one hand, the invention provides an embryo culture solution which contains sodium chloride, potassium chloride and other substances. On the other hand, the invention provides a method for cow in-vitro fertilization embryo culture. The method comprises the steps of 1 collecting oocytes and performing in-vitro maturation, 2 performing in-vitro fertilization and 3, performing embryo in-vitro culture and preservation. Germ cells are frozen and stored after being cultured in the embryo culture solution. The method has the excellent technological effect, for example, the germ cell development degree can be greatly increased.

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 in vitro fertilized embryos of the present invention has excellent technical effects, for example, it can obtain high culture efficiency with a low-cost method. In particular, the present invention relates to a method for in vitro fertilization and embryo culture of live or isolated bovine oocytes. More particularly, the presen...

Claims

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Application Information

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IPC IPC(8): C12N5/073C12N5/075
CPCC12N5/0604C12N5/0609C12N2500/12C12N2500/60C12N2501/02C12N2501/11C12N2501/31C12N2501/905
Inventor 王小武郭晶王娜郝少强赵明礼张月桥杨尚斌何伟马毅郭春明
Owner 天津力牧鼎丰生物技术有限公司
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