In-vitro culture solution for culturing swine parthenogenetic activated embryos and swine in-vitro fertilized embryos

A technology of embryo culture medium and in vitro fertilization, which is applied in the field of in vitro culture medium for cultivating porcine parthenogenetic activated embryos and in vitro fertilized embryos, can solve the problems of low blastocyst development rate and the number of cells, and achieve the effect of improving the efficiency of embryo development

Inactive Publication Date: 2014-04-09
BEIJING UNIV OF AGRI
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Problems solved by technology

[0010] To sum up, with the deepening of researchers' understanding of the environmental requirements of early embryo development and their own metabolic characteristics, the development rate and quality of in vitro embryos have been improved accordingly, but it still cannot fundamentally solve the problem of the existence of culture systems. Defects, blastocyst development rate and its small number of cells have not been well improved

Method used

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  • In-vitro culture solution for culturing swine parthenogenetic activated embryos and swine in-vitro fertilized embryos
  • In-vitro culture solution for culturing swine parthenogenetic activated embryos and swine in-vitro fertilized embryos
  • In-vitro culture solution for culturing swine parthenogenetic activated embryos and swine in-vitro fertilized embryos

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Experimental program
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Effect test

Embodiment 1

[0031] Embodiment 1, the effect of ligustrazine on the in vitro development of pig parthenogenetic activated embryos

[0032] One, the preparation of pig in vitro embryo culture medium containing ligustrazine

[0033] Take three NCSU-23 embryo culture fluids and add ligustrazine (Lig) to make the final concentrations reach 0.05 μg / mL, 0.5 μg / mL and 5 μg / mL respectively.

[0034] 2. Acquisition of porcine parthenogenetic activated embryos

[0035] 1. Oocyte collection and maturation culture

[0036] The pig ovaries collected from the slaughterhouse were stored in 34-37°C physiological saline supplemented with double antibodies (penicillin and streptomycin sulfate) and transported back to the laboratory within 1 hour. After the ovary was washed with the above normal saline three times, the surrounding fat and mesangium were removed, and the cumulus-oocyte complexes (COCs) in the follicles with a diameter of 2-6 mm on the surface of the ovary were extracted with a 12-gauge need...

Embodiment 2

[0077] Embodiment 2, Ligustrazine on the influence of pig IVF embryo in vitro development

[0078] One, the preparation of pig in vitro embryo culture medium containing ligustrazine

[0079] Take three NCSU-23 embryo culture fluids and add ligustrazine (Lig) to make the final concentrations reach 0.05 μg / mL, 0.5 μg / mL and 5 μg / mL respectively.

[0080] 2. Obtaining pig IVF embryos

[0081] 1. Oocyte collection and maturation culture

[0082] The pig ovaries collected from the slaughterhouse were stored in 34-37°C physiological saline supplemented with double antibodies (penicillin and streptomycin sulfate) and transported back to the laboratory within 1 hour. The ovary was rinsed with the above normal saline three times to remove the surrounding fat and mesangium, and the cumulus-oocyte complexes (COCs) in the follicles with a diameter of 2 to 6 mm on the surface of the ovary were extracted with a 12-gauge needle, and examined under a stereomicroscope. Select COCs with more...

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Abstract

The invention discloses a swine in-vitro embryo culture solution. The swine in-vitro embryo culture solution disclosed by the invention is obtained through adding ligustrazine, with the final concentration of 0.05-5.00 microgram/milliliter, into an embryo culture solution NCSU-23. A traditional Chinese medicine monomer component, namely ligustrazine, is added into the swine in-vitro embryo culture solution disclosed by the invention; shown by experiments, through carrying out in-vitro culture on the swine parthenogenetic activated embryos and the swine in-vitro fertilized embryos by using the culture solution, the development efficiency of the embryos and the number of the cell masses and total cells in blastulae can be effectively increased. The swine in-vitro embryo culture solution has the advantages that a foundation is laid for the development of other biotechnologies, medical research and animal husbandry, and meanwhile, a certain theoretical basis for clarifying the development mechanism of early swine embryos is provided.

Description

technical field [0001] The invention relates to a pig in vitro embryo culture solution, in particular to an in vitro culture solution for cultivating pig parthenogenetic activated embryos and in vitro fertilized embryos. Background technique [0002] The in vitro culture of animal embryos is an important link in embryonic biotechnology. Under in vitro culture conditions, early embryos often fail to complete the whole process of development from fertilized eggs to blastocysts, and stop at a specific developmental stage. This phenomenon is called in vitro block of embryonic development. , thus seriously affecting the quality of in vitro development of early embryos. For example, the development rate of pig blastocysts in vitro is generally only about 20%, and the number of inner cell masses is about 3, resulting in a decline in the birth rate of offspring, which greatly hinders the use of pigs in agriculture, bioengineering and Research applications in medicine. [0003] The...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073
Inventor 高建明史雅然赵菊李冬冬张志刚徐捷张超
Owner BEIJING UNIV OF AGRI
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