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Ovine oocyte in-vitro double-phase maturation method, pre-incubation liquid for ovine oocyte in-vitro double-phase maturation and kit

An oocyte and in vitro maturation technology, applied in the field of embryo engineering, can solve problems affecting the production efficiency of in vitro embryos, it is difficult to achieve simultaneous nucleoplasmic maturation, and the impact and application of sheep oocytes are unknown, so as to improve in vitro development ability, The effect of improving in vitro development ability and good application prospect

Active Publication Date: 2018-06-05
INNER MONGOLIA AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the effect and application of CNP to sheep oocytes remains unknown
[0005] Although conventional in vitro maturation of sheep oocytes can enable oocytes to complete meiosis in vitro, it is difficult to achieve synchronous maturation of nuclei and cytoplasm, which affects the efficiency of in vitro embryo production

Method used

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  • Ovine oocyte in-vitro double-phase maturation method, pre-incubation liquid for ovine oocyte in-vitro double-phase maturation and kit
  • Ovine oocyte in-vitro double-phase maturation method, pre-incubation liquid for ovine oocyte in-vitro double-phase maturation and kit
  • Ovine oocyte in-vitro double-phase maturation method, pre-incubation liquid for ovine oocyte in-vitro double-phase maturation and kit

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Effect test

experiment example

[0065] Experimental example, specific operation and effect verification of in vitro maturation of sheep oocyte of the present invention

[0066] 1. Mature culture of sheep oocytes

[0067] Sheep oocytes undergo conventional in vitro maturation and in vitro "two-stage" maturation provided by the present invention, the steps are as follows:

[0068] 1) Conventional in vitro maturation:

[0069] Step 1. Collection of oocytes: Use a sterile 10mL syringe 12 # The needle extracts follicles with a diameter of 3-6 mm from the surface of the ovary. Before collecting eggs, 1-2 mL of egg-collecting fluid should be extracted in a syringe, and cumulus oocytes with uniform cytoplasm and relatively complete cumulus cells should be selected under a stereo microscope. Cell complexes (Cumulus Oocyte Complexes, COCs) were used for in vitro maturation (In Vitro Maturation, IVM).

[0070] Step 2. Oocyte maturation culture: select the cumulus oocyte complex with uniform cytoplasm and relatively ...

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Abstract

The invention discloses an ovine oocyte in-vitro double-phase maturation method, pre-incubation liquid for ovine oocyte in-vitro double-phase maturation and a kit and belongs to the field of embryo engineering. The pre-incubation liquid is prepared from an oocyte in-vitro basic culture medium TCM199 added with C-type natriuretic peptide and polyvinylpyrrolidone (PVP). Therefore, the C-type natriuretic peptide provided by the invention is pre-incubated and is combined with a double-phase culture system of in-vitro maturation, so that nucleoplasm synchronous maturation of an ovine oocyte can beeffectively facilitated and the in-vitro development capability of the oocyte is improved; high-quality ovum sources and embryos can be provided for ovine in-vitro embryo production; requirements on scientific researches and production are met better.

Description

technical field [0001] The invention relates to the field of embryo engineering, in particular to a method for in vitro "two-stage" maturation of sheep oocytes, a pre-incubation solution and a kit. Background technique [0002] The quality of in vitro maturation of sheep oocytes is a key factor affecting in vitro fertilization and in vitro embryo development, and it is also the dominant factor restricting the source of embryo transfer, embryonic stem cells and transgenic animals. breakthrough. Oocyte maturation in vitro goes through a multi-stage, complex, and discontinuous process involving nuclear maturation, cytoplasmic maturation, and the interaction between granulosa cells and oocytes. [0003] For a long time, researchers have been trying to simulate the physiological internal environment of oocyte survival and development, improve the in vitro maturation system of oocytes, provide high-quality egg sources for production and scientific research, and then improve the e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/075
CPCC12N5/0609C12N2500/50C12N2501/30
Inventor 张通张家新海日汗王兆琛
Owner INNER MONGOLIA AGRICULTURAL UNIVERSITY
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