Buffalo oocyte in-vitro maturation (IVM) culture method

A technology of in vitro maturation culture and oocyte, applied in the field of in vitro maturation and culture of buffalo oocytes, can solve the problems of low development rate of buffalo oocytes, etc. Effect

Inactive Publication Date: 2016-06-01
云南中科胚胎工程生物技术有限公司
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  • Abstract
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  • Application Information

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Problems solved by technology

[0004] Aiming at the problem of low developmental rate of buffalo oocytes after fertilization by existi

Method used

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Examples

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Effect test

Embodiment 1

[0024] Example 1: Step 1. Preparation of oocytes: Collect buffalo ovaries, and transport all ovaries in a thermos flask at 25-35°C in physiological saline containing double antibodies within 3 to 4 hours. Wash your hands with soap, put the ovaries in a wide-mouth thermos, pour the anti-biological saline solution to submerge the ovaries, wash the ovaries with your hands, wash the blood on the ovaries as much as possible, dry them with sterile gauze, and suck 2mm in diameter on the ovaries If there are antral follicles, put the follicular fluid together with the oocyte complexes (COCs) into the test tube (the test tube is placed in a constant temperature water bath at 30~35℃).

[0025] Step two, IVM culture droplet preparation: prepare IVM droplets at least 2h before aspirating oocytes. The preparation of IVM droplets must be performed on a clean bench. Use a micropipette to suck up 50μLIVM culture solution, drop it into a 35mm diameter petri dish, prepare 7 IVM droplets per petri...

Embodiment 2

[0028] Example 2: Step 1. Preparation of oocytes: Collect buffalo ovaries, and transport all ovaries in a thermos flask at 25-35°C in physiological saline containing double antibodies within 3 to 4 hours. Wash your hands with soap, put the ovaries in a wide-mouth thermos, pour the anti-biological saline solution to submerge the ovaries, wash the ovaries with your hands, wash the blood on the ovaries as much as possible, and absorb 8mm in diameter on the ovaries. If there are antral follicles, put the follicular fluid together with the oocyte complexes (COCs) into the test tube (the test tube is placed in a constant temperature water bath at 30~35℃).

[0029] Step two, IVM culture droplet preparation: prepare IVM droplets at least 2h before aspirating oocytes. The preparation of IVM droplets must be performed on a clean bench. Use a micropipette to suck up 50μLIVM culture solution, drop it into a 35mm diameter petri dish, prepare 7 IVM droplets per petri dish. After the droplets...

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Abstract

The invention relates to a buffalo oocyte in-vitro maturation (IVM) culture method. The method is characterized by comprising the following steps: oocyte preparation, IVM culture droplet preparation and in-vitro maturation culture. The IVM culture fluid is composed of TCM199, FBS, oFSH, 17bet-E2 and GSH. The buffalo oocyte IVM culture method can obviously promote the maturation of oocyte cytoplasms, and solves the problem of low embryo ectogenesis rate after the buffalo in-vitro mature oocytes are fertilized. The maturation culture fluid can promote the maturation of the buffalo oocytes, and has favorable effects on maintaining the morphology of the oocytes and early embryos.

Description

Technical field [0001] The invention belongs to the technical field of embryo engineering, in particular to a method for in vitro maturation and culture of buffalo oocytes. Background technique [0002] As the basis of embryo bioengineering research, the only in vitro maturation and culture technology of bovine oocytes. Since the 1930s, many scientific researchers have done a lot of work in the field of bovine oocyte growth and development in vitro. The cultivation conditions have been studied extensively and in-depth, and great progress has been made. [0003] In recent years, great progress has been made in the in vitro maturation technology of buffalo oocytes, and a more mature in vitro maturation technology system has been formed. However, there are still many gaps compared with in vivo maturation, especially the rate of blastocysts and blastocysts. The low quality results in low development rate of buffalo oocytes after fertilization. The existing in vitro maturation culture...

Claims

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Application Information

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IPC IPC(8): C12N5/075
CPCC12N5/0609C12N2501/30C12N2501/31C12N2501/999
Inventor 苏雷
Owner 云南中科胚胎工程生物技术有限公司
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