Bovine in vitro fertilization embryo culture medium and culture method

A technology of embryo culture solution and culture method, which is applied in the field of agriculture-animal husbandry and veterinary medicine, can solve the problems of high production cost, low efficiency, and limited wide application of in vitro fertilized embryos, and achieve the goals of accelerating the genetic breeding process, reducing costs, and improving developmental ability Effect

Inactive Publication Date: 2016-04-20
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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  • Abstract
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Problems solved by technology

[0007] 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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  • Bovine in vitro fertilization embryo culture medium and culture method
  • Bovine in vitro fertilization embryo culture medium and culture method

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Embodiment Construction

[0020] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are all commercially available products.

[0021] The culture method of embodiment bovine in vitro fertilization embryo

[0022] 1. Reagents

[0023] Mature medium: TCM-199 containing 0.01IU / mlFSH, 10IU / mlLH, 1μg / ml estradiol, 100ng / mlIGF, 50ng / mlEGF, 100U / ml penicillin, 100μg / ml streptomycin, 10% fetal bovine serum culture medium.

[0024] Semen washing liquid: containing 112.0mM NaCl, 4.02mMKCl, 2.25mM CaCl 2 2H 2 O, 0.52mMMgCl 2 ·6H 2 O, 0.83mMKH 2 PO 3 , 37.0mM NaHCO 3 , 1.25 mM sodium pyruvate, 10 μg / ml heparin, 4 mg / ml bovine serum albumin (Bovine serum albumin, BSA), 10 mM caffeine, 100 U / ml penicillin, 100 μg / ml streptomycin in water.

[0025] Semen fluid: contain...

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Abstract

The invention provides a culture fluid exclusively used in cow in-vitro fertilization embryos. The culture fluid contains 109.0mM-110mM of NaCl, 2.9mM-3.1mM of KCl, 26.0mM-26.5mM of NaHCO3, 0.5mM-1.0mM of MgCl2.6H2O, 1.0mM-1.3mM of KH2PO3, 0.4mM of sodium pyruvate, 1.5 mM of glucose, 5mM of galacturonic calcium, 10v / v% fetal bovine serum, 1mM of L- glutamine, 2v / v% essential amino-acid, 1v / v% nonessential amino acid and 1mM-10mM of glutathione. The cow in-vitro fertilization embryos are placed into the culture fluid to carry out in-vitro cultivation, and results are shown to be obviously superior to those of a control group which is not added with GSH (glutathione), and therefore, developmental rate of blastula and embryo quality are improved, cost for producing embryos in vitro is lowered, experimental basis is provided for applying a cow IVF (in vitro fertilization) technology to practice, and a genetic breeding process can be greatly accelerated.

Description

technical field [0001] The invention belongs to the field of agriculture-animal husbandry and veterinary medicine, and in particular relates to a bovine in vitro fertilized embryo culture solution and a culture method. Background technique [0002] With the development of modern agricultural science and technology, in order to make full use of the reproductive potential of fine-bred cows and accelerate the process of genetic breeding, it is inevitable to apply efficient new breeding technologies in production practice. OvumpickUP (OPU) and in vitro fertilization (InVitroFertilization, IVF) are new technologies for embryo engineering that developed rapidly in the 1980s. The combination of the two can obtain a large number of embryos with a clear genetic lineage, thereby shortening the generation interval. . At present, these two technologies have become important breeding technologies adopted by farmers in developed animal husbandry countries such as Europe, America and Ocea...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/073
Inventor 杜卫华孙尉俊朱化彬
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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