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Porcine oocyte in vitro maturation medium and methods of preparation and culture

A technology for in vitro maturation culture and oocyte, which is applied in the field of culturing pig oocytes, in vitro maturation medium of pig oocytes and their preparation, which can solve the problems of low maturation rate and achieve the effects of improving maturation rate and simple operation.

Inactive Publication Date: 2016-04-20
THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

However, these porcine oocytes cultured in vitro show that the maturation rate is very low, and we need to improve the in vitro culture medium to increase the maturation rate of porcine oocytes cultured in vitro

Method used

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  • Porcine oocyte in vitro maturation medium and methods of preparation and culture
  • Porcine oocyte in vitro maturation medium and methods of preparation and culture
  • Porcine oocyte in vitro maturation medium and methods of preparation and culture

Examples

Experimental program
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Embodiment 1

[0040] The formulation of the balanced oocyte in vitro culture solution (invitromature, IVM solution) used in the present embodiment is as follows:

[0041]

[0042] Balance of culture solution: Add 1mL of the above-mentioned IVM solution to a 60cm glass petri dish to make it cover the bottom, then cover the surface of the liquid with 500μL of mineral oil, and place in 5% CO 2 The incubator with saturated humidity was equilibrated for at least 6 hours.

[0043] Oocyte collection: Use a 10mL syringe to provide negative pressure to extract eggs, select 4-7mm follicles, and insert the needle in parallel. During the collection process, it is necessary to avoid inserting into the medulla, and extract too many tissue fragments, which will make it difficult to collect eggs and delay the time of egg collection. When picking oocytes, use a simple egg picking device. One end of the silicone tube is tightly covered with an egg picking needle, and the other end is tightly set with a 1...

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Abstract

The invention discloses a porcine oocyte in vitro maturation medium and methods of preparation and culture. The medium comprises a TCM-199 base medium and the added porcine follicular fluid, fetal bovine serum, eCG, hCG, EGF and IGF-1. The porcine oocyte in vitro maturation medium of the invention can significantly increase the rate of maturation of porcine oocytes in vitro. Experiments confirm that after the oocyte collection and 42-44h maturation, the medium of the invention can have a polar body rate stabilized at 80% and a parthenogenetic embryo development rate up to 85%, and the entire maturation process does not require replacement of the medium and is easy to operate.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a porcine oocyte in vitro maturation culture medium and a preparation method thereof, and a method for cultivating porcine oocytes using the culture medium. Background technique [0002] In vitro culture and maturation of oocytes is the basis and key link of embryonic biotechnology. At present, embryo technology has greatly promoted the progress of animal husbandry and medicine, such as breed improvement, breeding of transgenic disease-resistant varieties, preparation of human disease models, production of medicinal proteins in animal bioreactors, and exploration of developmental mechanisms. [0003] Somatic cell cloning of pigs and preparation of disease models require a large number of oocytes with developmental potential, but the current in vitro maturation rate of pig oocytes is not ideal. [0004] At present, there are the following in vitro culture base fluids for pig...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/075
Inventor 蔡志明牟丽莎冯万有
Owner THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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