Bovine oocyte in vitro maturation serum-free medium

A serum-free medium and in vitro maturation technology, applied in the field of cell biology, can solve problems such as difficult to rule out the interference of growth factors in serum

Inactive Publication Date: 2008-07-23
BEIJING JINXIU DADI AGRI CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In addition, in some specific experimental situations, if it is necessary to observe the effect of a growth factor on a certain cell, it is difficult to rule out the interference of the growth factor in the serum.

Method used

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Embodiment Construction

[0027] The present invention is described in further detail below by embodiment.

[0028] 1. Main equipment

[0029] CO 2 Incubator, constant temperature table, stereo microscope, water bath, constant temperature box, ultra-clean table, balance, volumetric flask.

[0030] 2. In Vitro Maturation of Bovine Oocytes

[0031] Collect the ovaries from the slaughterhouse. Use 25°C normal saline with double antibody and transport it to the laboratory within 3 hours. Use a syringe to extract follicles with a size of 2-7 mm on the ovary, and select oocyte complexes (COCs) with several layers of granulosa cells and uniform cytoplasm under a stereoscopic anatomical microscope for in vitro maturation and culture, and mature in vitro for 24 hours , to expel polar bodies as a sign of maturity.

[0032]IVMBM-I culture medium composition: TCM199+10μg / ml FSH (Follicle-stimulating Hormone)+1μg / mlLH (Luteinizing Hormone)+1μg / ml Estradiol+0.0625g Penicillium+0.05g Streptomycin, added ingredie...

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Abstract

The invention relates to a serum free culture medium for in vitro maturation of bovine oocyte, belonging to the field of cytobiology, in particular to a serum free medium IVMBM-I for in vitro maturation of bovine oocyte, comprising a TCM199 medium, hormone for oocyte maturation and antibacterial agent, which is characterized in that: Sodium pyruvate, Taurine, Insulin, Selenium, BSA, HEPES, TGF-a, L-Glutamine and EGF are added, and the serum is not contained. The serum free culture medium for in vitro maturation of bovine oocyte has the advantages of avoiding the adverse effect of some components in the serum on the oocyte, exact quantification and good repeatability because the conventional serum is substituted by the added components, and that the function of the serum in the maturation process of the oocyte can be completely substituted by the added components is proved by the experiments of in vitro maturation, in vitro fertilization and in vitro development.

Description

technical field [0001] The invention belongs to the field of cell biology and relates to a serum-free medium for in vitro maturation of bovine oocytes. Background technique [0002] Animal cell culture is the most widely used in vitro culture technology, which originated in the 19th century. In 1885, scientists successfully maintained the neural plates of chicken embryos in warm salt water for several days, which was the first attempt to culture them in vitro. With the development of antibiotics, synthetic culture medium, trypsin treatment technology and the improvement of vessels, animal cell culture technology has been developed rapidly. The selection and preparation of culture medium is a key link in in vitro culture technology. The general culture medium consists of two parts: basal medium and serum. [0003] Basal medium Self-prepared or ready-made chemically synthesized medium, commonly used chemically synthesized medium includes Eagle (including BME, MEM, DME, etc....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N5/075
Inventor 王海张家新张红霞安晶张向利吴亦芳赵孟彬王玉忠
Owner BEIJING JINXIU DADI AGRI CO LTD
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