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Two-stage culture method of bovine embryo in vitro

A technology of two-stage embryo culture and embryo culture, which is applied in the field of applied embryo biology, and can solve the problems of poisoning, unfavorable development of bovine embryos, glucose reducing blastocyst rate, etc.

Inactive Publication Date: 2013-05-15
INNER MONGOLIA UNIV FOR THE NATITIES
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AI Technical Summary

Problems solved by technology

Although glucose can be utilized during the development of bovine implanted embryos, many studies have found that the addition of glucose to SOF solution is toxic to early embryo development, and some studies have shown that glucose reduces the blastocyst rate during in vitro culture.
However, if glucose is not added or the concentration of glucose is too low, it is also not conducive to the development of bovine embryos.

Method used

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  • Two-stage culture method of bovine embryo in vitro

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Embodiment 1

[0017] Example 1 Bovine Embryo Culture in Vitro

[0018] The invention establishes a novel in vitro culture system for bovine embryos by adopting two kinds of SOF culture solutions containing different concentrations of glucose to cultivate bovine embryos. Two-stage culture method is mainly adopted: firstly, the embryos are cultured in SOF solution containing 0.15mM concentration for 5 days, then cultured in SOF solution containing 2mM concentration for 2 days, and the blastocyst rate is counted. The SOF solution of glucose was cultured for the first stage as a control, and the blastocyst rate was counted after 7 days of culture.

[0019] Specifically, the bovine embryo in vitro culture program using SOF solution of the present invention:

[0020] (1) In vitro maturation of oocytes: Wash the selected grade A and B cumulus-oocyte complexes (COCs) three times in egg wash solution (TCM199-Hepes), and mature solution (containing FSH10μg / ml, LH1μg / ml, E21μg / ml, EGF10ng / ml, TCM19...

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Abstract

The invention discloses a two-stage culture method of a bovine embryo in vitro. The method comprises the steps of culturing a bovine oosperm for 2-3 days in an SOF (Synthetic Oviduct Fluid) containing 0.12-0.16mM of glucose, and then culturing for 4-5 days in the SOF containing 1.8-2.2mM of the glucose. The invention also provides a reagent for culturing the bovine embryo in vitro. The reagent comprises a solution A, namely the SOF containing 0.12-0.16mM of the glucose, and a solution B, namely the SOF containing 1.8-2.2mM of the glucose. According to the invention, the blastocyst development rate is increased by regulating the glucose concentration and the culture time based on a traditional SOF, and the SOF provided by the invention can be used as an in-vitro embryo culture solution to be popularized and applied in the livestock production.

Description

technical field [0001] The present invention relates to a novel in vitro embryo culture method, in particular to the application of synthetic oviduct fluid (SOF) in the in vitro culture of bovine embryos, which can be used to produce transgenic clones using oocytes obtained from superovulated or abattoir ovaries Livestock, livestock breeding and protection of endangered species. The invention belongs to the field of applied embryo biotechnology. Background technique [0002] In vitro embryo culture technology is an important link in embryo transfer technology. In vitro embryo culture is carried out after in vitro fertilization of ovarian oocytes derived from superovulation or abattoirs, so as to give full play to the reproductive potential of excellent female animals and provide sufficient embryo sources for embryo transfer. The application of this technology doubles the number of good offspring, and accelerates the pace of multiplication of good varieties and animal breed...

Claims

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Application Information

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IPC IPC(8): C12N5/073
Inventor 贾振伟张显华张颖
Owner INNER MONGOLIA UNIV FOR THE NATITIES
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