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Culture medium for improving heat resistance of embryos and method of using the same

An embryo culture and culture medium technology, applied in the fields of embryo engineering and developmental biology, can solve problems such as no obvious effect, and achieve the effects of outstanding effect, simple preparation method and reduced economic cost.

Active Publication Date: 2017-05-24
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There was no apparent effect on the thermotolerance of embryos

Method used

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  • Culture medium for improving heat resistance of embryos and method of using the same
  • Culture medium for improving heat resistance of embryos and method of using the same
  • Culture medium for improving heat resistance of embryos and method of using the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Heat-resistant culture experiment of bovine embryos

[0025] (1) Prepare heat-resistant culture medium suitable for bovine embryos

[0026] Table 1 Composition of conventional SOF medium for bovine embryos

[0027] ingredient mg / 100ml NaCl629 KCl53 CaCl 2 ·2H 2 O

25 MgCl 2 ·6H 2 O

10 KH 2 PO 4

16 NaHCO 3

211 Sodium lactate60ul Sodium pyruvate3.6 glucose2.7 phenol red0.02ml PVA 1mg / ml

[0028] The above reagents were purchased from sigma company.

[0029] First, prepare 100ml of conventional SOF culture medium (see Table 1 for details). Measure 90ml of ultrapure water into a sterilized beaker, accurately weigh the various components in Table 1, and add them to the beaker containing ultrapure water to obtain a conventional SOF culture solution. After each ingredient is added, stir and shake with a magnetic stirrer until it is completely dissolved before adding the next ingredient.

[0030] Accurately weigh 8mg of baicalin, first dissolve it with 10mL of...

Embodiment 2

[0047] Different from Example 1, when preparing a heat-resistant culture medium suitable for bovine embryos, the amount of baicalin is 2 mg and the amount of glutamine is 117 mg.

Embodiment 3

[0049] Different from Example 1, when preparing a heat-resistant culture solution suitable for bovine embryos, the amount of baicalin is 10 mg and the amount of glutamine is 175 mg.

[0050] Table 2 The influence of the culture solution of the present invention on the development rate of early bovine embryos at different temperatures

[0051]

[0052] Note: Cleavage rate = number of divided embryos / total fertilized eggs × 100%; morula rate = number of morula embryos / number of split embryos × 100%; blastocyst rate = number of blastocysts / number of split embryos × 100%; hatching blastocysts Rate=number of hatched blastocysts / number of split embryos×100%; the same letter in the table indicates that the difference is not significant (P> 0.05); different letters indicate significant differences (P <0.05).

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Abstract

According to the difficulty in embryo heat resistance in the prior art, the invention provides an embryo heat resistance improving culture solution. The embryo heat resistance improving culture solution takes an ordinary culture solution as a substrate, wherein the ordinary culture solution is a culture solution for embryo culture; the embryo heat resistance improving culture solution further comprises a baicalin and a glutamine; the concentration of the baicalin is 2.0-10.0 micrograms / mL, and the concentration of the glutamine is 1.17-1.75mg / ml. A heat-resistant culture solution is prepared by adding the baicalin and the glutamine at proper concentrations into the embryo culture solution, so that not only the economic cost is reduced and the preparation method is simple, but also the rate of embryonic development during heat stress and the blastocyst hatching rate are substantially improved; the baicalin and the glutamine have a synergistic effect and both can adjust expression of HSP70 (heat shock protein 70), so that compared with the single baicalin or glutamine, the prepared heat-resistant culture solution has a prominent effect and can generate an unexpected technical effect.

Description

Technical field [0001] The invention belongs to the field of embryo engineering and developmental biology, and specifically relates to a culture medium for embryos, in particular to a culture medium for improving the heat resistance of early embryos and a method of use. Background technique [0002] As the global warming and temperature rise, the impact of high temperature on humans and animals becomes more and more obvious. High temperature in summer and certain heat (increasing body temperature) diseases will cause heat stress in animals and lead to the death of pregnant animal embryos. As a result, its fecundity is reduced or even infertility, which causes great economic losses to animal husbandry production. [0003] The traditional culture medium is formulated to simulate the environment when the embryo develops in the fallopian tube, and its main components include: water, ions, energy substances, amino acids, vitamins, nucleic acid precursors, proteins, growth factor hormone...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/073
Inventor 田文儒赵园丛霞曹荣峰李华涛姜忠玲崔凯齐晓楠
Owner QINGDAO AGRI UNIV