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A kind of in vitro culture medium and culture method for porcine parthenogenetic activated embryo

An in vitro culture and parthenogenetic activation technology, which is applied in the direction of culture process, tissue culture, cell culture active agent, etc., can solve the problems of insufficient quality and effect of embryo culture, and the inability to ensure the stability of oocyte activation, etc., to achieve Guaranteed chromosomal multiples, beneficial to cell division, and increased flexibility

Active Publication Date: 2020-06-16
SHANGHAI ACAD OF AGRI SCI
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the existing in vitro culture medium and culture methods for porcine parthenogenetic activation of embryos cannot guarantee the stability of oocyte activation, cannot provide the energy required for cell metabolism, and are not enough to obtain high embryo culture quality and effects

Method used

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  • A kind of in vitro culture medium and culture method for porcine parthenogenetic activated embryo
  • A kind of in vitro culture medium and culture method for porcine parthenogenetic activated embryo
  • A kind of in vitro culture medium and culture method for porcine parthenogenetic activated embryo

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Embodiment

[0030] 1. Preparation of porcine oocyte maturation culture medium

[0031] In vitro maturation culture medium is composed of TCM-199 plus 10% (V / V) fetal bovine serum (Gibco, the United States), 10% (V / V) porcine follicular fluid (made in the laboratory), 10IU / mL pregnant horse serum gonadotropin (Ningbo Third Hormone Products Co., Ltd., China), 10IU / mL human chorionic gonadotropin (Ningbo Third Hormone Products Co., Ltd., China), 10IU / mL double antibody (Sigma, USA), 0.1mg / mL L-cysteine ​​(Sigma, USA) and 10ng / mL epidermal growth factor (Sigma, USA) were mixed.

[0032] 2. Acquisition and in vitro maturation of porcine oocytes

[0033] Pig ovaries were collected from the slaughterhouse, placed in 37°C normal saline (containing 1000 IU / mL penicillin and streptomycin), and transported back to the laboratory within 2 hours. Use a 18-gauge needle syringe to extract cumulus-oocyte complexes (COCs) with a follicle diameter of 2-8 mm and inject them into a 15-mL centrifuge tube. A...

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Abstract

The present invention discloses an in vitro culture liquid (mPZM-3) and a culture method for a pig parthenogenetic activation embryo, and belongs to the field of in vitro animal embryo culture. The in vitro culture liquid (mPZM-3) comprises raw materials such as a mPZM-3 base culture liquid, calcium lactate pentahydrate and sodium pyruvate, wherein the concentration of the calcium lactate pentahydrate in the in vitro culture liquid is 0.606 g / L, the concentration of the sodium pyruvate is 0.022 g / L, the solvent used by the in vitro culture liquid is hydrogen-rich water, and the H2 content is 1.2-1.6 ppm. The culture method for the pig parthenogenetic activation embryo by using the in vitro culture liquid comprises: washing a pig parthenogenetic activation embryo by using an in vitro culture liquid containing 5 [mu]mol / L cytochalasin B, treating for 3-5 h in the in vitro culture liquid containing 5 [mu]mol / L cytochalasin B, washing with a cytochalasin B-free in vitro culture liquid, and culturing for 168 h in the cytochalasin B-free in vitro culture liquid until the embryo develops into a blastula. According to the present invention, with the in vitro culture liquid and the culture method, the energy and the development environment can be well provided for the ig parthenogenetic activation embryo so as to improve the culture quality and the culture effect of the embryo.

Description

technical field [0001] The invention relates to the field of in vitro culture of animal embryos, in particular to an in vitro culture solution and a culture method for pig parthenogenetic activated 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 30, 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 in vitro cul...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/073
CPCC12N5/0604C12N2500/02C12N2500/12C12N2500/16C12N2500/30C12N2500/32C12N2500/33C12N2501/30
Inventor 张德福戴建军张树山吴彩凤陈亚宁王文杰
Owner SHANGHAI ACAD OF AGRI SCI
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