Efficient vitrification freezing method for bovine in-vitro embryo production

A vitrification, bovine body technology, applied in biochemical equipment and methods, cell dissociation methods, preservation of human or animal bodies, etc., can solve the problems of limited application, low pregnancy rate of transplantation, etc. The effect of efficiency

Pending Publication Date: 2022-05-27
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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Problems solved by technology

However, the pregnancy rate of bovine in vitro embryo production after freezing (35.89+3.87%) was significantly lower

Method used

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  • Efficient vitrification freezing method for bovine in-vitro embryo production
  • Efficient vitrification freezing method for bovine in-vitro embryo production
  • Efficient vitrification freezing method for bovine in-vitro embryo production

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Embodiment 1

[0038] Example 1 Efficient vitrification method for bovine in vitro embryo production

[0039] 1. Experimental method

[0040] 1. In vitro maturation of oocytes

[0041] The cumulus-oocyte complexes (COCs) in follicles with a diameter of 2-8 mm were collected from bovine ovaries (collected from Dachang, Hebei), and COCs containing three or more layers of granulosa cells were selected for in vitro maturation solution (IVM solution). : TCM199+10μg / mL FSH+10μg / mL LH+10μg / mL E2+10%FBS) after washing 3 times, put 50 COCs / well / 500μL mature medium into the pre-equilibrated four-well plate culture medium for 2h , at 38.5℃, 5%CO 2 and cultured in a saturated humidified carbon dioxide incubator.

[0042] 2. Production of in vitro fertilized embryos

[0043] Embryos produced by bovine in vitro fertilization refer to the method of Nedambale et al., with slight modifications.

[0044] After COCs were cultured for 22-24 h in vitro, granulosa cells were removed by digestion with 0.1% hy...

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Abstract

The invention provides an efficient vitrification freezing method for bovine in-vitro embryo production. According to the method, beta-nicotinamide mononucleotide is applied to a bovine IVF embryo vitrification technology in bovine in-vitro embryo production for the first time, the beta-nicotinamide mononucleotide with a certain concentration is added into a bovine oocyte in-vitro maturation solution and a bovine in-vitro fertilization embryo culture solution, and then the bovine in-vitro embryo is subjected to vitrification. The vitrification efficiency of the bovine IVF embryo can be greatly improved. The vitrification efficiency can be further improved by further adding growth factors and gap-linked proteins into a bovine oocyte in-vitro maturation solution and a bovine in-vitro fertilized embryo culture solution according to a proportion. Furthermore, by adding the nanoparticles with a certain concentration into the vitrification freezing liquid, the vitrification freezing efficiency can be further improved.

Description

technical field [0001] The invention relates to the technical field of bovine in vitro embryo production, in particular to a high-efficiency vitrification and freezing method for bovine in vitro embryo production. Background technique [0002] The production of bovine in vitro embryos has important economic and social value. It can be used to provide embryo sources for cloning, nuclear transfer, and transgenic animal production, rapidly multiply excellent livestock groups, and protect endangered animals as a means of germplasm resource preservation. It is of great importance theoretical and social significance. However, the post-frozen transfer pregnancy rate of bovine in vitro embryo production (35.89+3.87%) was significantly lower than that of fresh in vitro embryo production (51.35+1.87%), which greatly limited their application (Niu et al., 2014). According to IETS statistics, from 1997 to 2017, the global bovine in vitro embryo transfer was dominated by fresh embryo tr...

Claims

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Application Information

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IPC IPC(8): C12N5/075A01N1/02
CPCC12N5/0609A01N1/0221A01N1/021A01N1/0226C12N2500/02C12N2501/31C12N2500/35C12N2500/40C12N2509/00C12N2509/10
Inventor 赵学明徐茜
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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