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Culture solution used for embryo vitro production and method for bovine embryo vitro production

A culture medium and embryo body technology, applied in the biological field, can solve the problems of reducing the efficiency of embryo development into blastocyst, unable to fully provide early embryo nutrition and non-nutritive substances, restricting the development of animal husbandry, etc., to improve quality and efficiency, improve Effects of blastocyst development rate

Inactive Publication Date: 2010-11-17
周虚
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, pure basic culture medium such as TCM-199, CZB, KSOM, etc. cannot completely simulate the environment of embryonic development in vivo, and cannot fully provide the nutrients and non-nutrient substances necessary for early embryonic development, thus greatly reducing the risk of embryonic development into blastocysts. s efficiency
As far as the in vitro production of early bovine embryos is concerned, these problems are particularly prominent, which greatly restricts the development of animal husbandry

Method used

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  • Culture solution used for embryo vitro production and method for bovine embryo vitro production
  • Culture solution used for embryo vitro production and method for bovine embryo vitro production
  • Culture solution used for embryo vitro production and method for bovine embryo vitro production

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Detection of BDNF mRNA expression level in early embryos

[0034] The expression levels of BDNF in oocytes and embryos at different developmental stages and early embryos produced by different methods (in vitro fertilization embryos, parthenogenetic activation, nuclear transfer embryos) were detected by fluorescent quantitative PCR technology. The results are shown in Table 1 and Table 2.

[0035] Table 1 Expression levels of BDNF mRNA in bovine oocytes and in vitro fertilized embryos at different developmental stages

[0036]

[0037] Note: Different capital letters indicate extremely significant difference (P<0.01)

[0038] Table 2 BDNF mRNA expression levels in different types of bovine embryos

[0039]

[0040] Note: Different capital letters indicate extremely significant difference (P<0.01)

[0041] The above results indicated that BDNF was expressed in oocytes and embryos at different developmental stages and early embryos produced by different m...

Embodiment 2

[0042] Example 2: Detection of BDNF protein expression in embryos by immunofluorescence

[0043] The expression of BDNF protein in embryos was detected by immunofluorescence technique, which showed that the signal of BDNF protein was mainly concentrated in the ectoderm of blastocysts.

Embodiment 3

[0044] Example 3: Collection and in vitro maturation of oocytes

[0045] Bovine ovaries collected from the slaughterhouse were placed in a K + , Ca 2+ and Mg 2+ Put it in a 35-37°C normal saline thermos, send it to the laboratory within 4 hours, and wash it with 37°C normal saline. Then use a 10 mL syringe with a 12-gauge needle to extract oocytes from follicles with a diameter of 2-6 mm. Select oocytes with uniform cytoplasm and complete or partially dense cumulus cells under a solid microscope, wash them twice with egg washing solution, and put them into a glass plate (30×10mm) containing 1.5mL mature culture medium. At 38.5°C, 5% CO 2 and cultured in an incubator with saturated humidity for 22-24 hours to obtain mature oocytes.

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Abstract

The invention relates to the technical field of biology and discloses culture solution used for embryo vitro production. The culture solution is TCM-199 culture solution comprising 5 to 10mmol / L of Hepes, 15 to 26.2mmol / L of NaHCO3, 30 to 50mu g / L of brain derived neurotrophic factor (BNDF) and 3 to 5 percent OCS, wherein the concentration of the BDNF in the culture solution is preferably 40mu g / L. The culture solution of the invention can obviously improve the ratio of blastocyst developed from bovine embryo body. The invention also provides a method for performing bovine embryo vitro production by using the culture solution.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a culture solution for in vitro production of embryos and a method for in vitro production of bovine early embryos. Background technique [0002] Animal embryo transfer refers to the transfer of fertilized eggs or early embryos that have developed to a certain period from the reproductive tract of a pregnant female animal (donor) to a certain part of the reproductive tract of a female animal (recipient) that is in estrus synchronously with the donor female animal. , to make it continue to grow, develop, and grow into offspring technology. The use of embryo transfer can shorten the animal reproduction cycle, further excavate the animal's reproductive potential, provide an effective solution for the mass reproduction of high-quality livestock and the continuation of rare animals, and play an important role in the fields of animal husbandry and pharmaceutical industry. [0003] Since t...

Claims

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Application Information

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IPC IPC(8): C12N5/073
Inventor 周虚李纯锦易康乐
Owner 周虚
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