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Method for improving in-vitro production efficiency of buffalo embryos

A technology of production efficiency and water buffalo, applied in the field of bioengineering, can solve problems such as unsatisfactory, and achieve the effect of improving production efficiency

Inactive Publication Date: 2011-08-03
GUANGXI UNIV
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  • Description
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AI Technical Summary

Problems solved by technology

But judging from the current situation of my country's milk buffaloes, their quantity and milk production are far from meeting the market demand.

Method used

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  • Method for improving in-vitro production efficiency of buffalo embryos
  • Method for improving in-vitro production efficiency of buffalo embryos
  • Method for improving in-vitro production efficiency of buffalo embryos

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Embodiment Construction

[0016] Buffalo ovaries were collected from the slaughterhouse, placed in a vacuum flask with normal saline at 30-35°C and sent back to the laboratory within 4 hours. After the ovaries were sent back to the laboratory, the ovaries were washed 3 to 4 times with sterile saline. Then use a disposable 10 mL syringe equipped with a 18-gauge needle to extract oocytes from follicles with a diameter of 2 to 8 mm on the surface of the ovary. Under a stereo microscope, select oocytes wrapped with three or more layers of dense cumulus cells, with bright appearance and uniform cytoplasm, wash the oocytes with the collection solution for 2 to 3 times, and then wash them with the maturation solution for 2 times. Then put it into a glass Petri dish (20×10mm) containing 1mL of mature culture medium, at 38.5°C, 5% CO 2 In vitro maturation culture was carried out in an incubator with maximum saturated humidity for 22-24 hours.

[0017] The composition of oocyte collection fluid is:

[0018] T...

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Abstract

The invention relates to a method for improving in-vitro production efficiency of buffalo embryos, comprising the following steps of: respectively adding 10 muL / mL of insulin-transferrin-selenium (ITS) and 20 muL / mL of insulin-transferrin-selenium (ITS) to an oocyte maturing solution and a culture solution, and then carrying out in-vitro maturation and in-vitro culture. The method indicates that the in-vitro maturation rate of buffalo oocyte is effectively improved by adding 10 muL / mL ITS to the maturing solution, and the exhaustion ratio of a first polar body is up to 65.61%; and adding 20 muL / mL of ITS to the embryo culture solution is beneficial to the growth of early embryos of a fertilized buffalo, and the growing rate of a blastaea is up to 29.93%. the method has the advantages of improving the in-vitro maturation rate of the buffalo oocyte and the growing rate of the blastaea, thereby improving the in-vitro production efficiency of the buffalo embryos and providing a large number of high-quality embryos for embryo transplantation.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, in particular to a method for producing mammalian embryos in vitro. Background technique [0002] Insulin-transferrin-selenium (insulin, transferrin and selenium, ITS) is mainly composed of insulin, transferrin and selenium. Insulin stimulates the synthesis of DNA, RNA, proteins and lipids, promotes the utilization of glucose and amino acids, and regulates cellular functions through its effects on the plasma membrane, cytoskeleton, intracellular enzymes and nucleus. At the same time, insulin also promotes cell mitosis. Transferrin is a serum globulin, which is related to the transport of iron. When the receptor on the cell surface binds to transferrin, iron can be transferred into the cell; and the cell participates in O 2 and CO 2 The metabolic and cellular respiration processes of myoglobin and hemoglobin synthesis are mainly dependent on protein and iron. In addition, transferrin ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073
Inventor 张明陈焕华付强黄怡
Owner GUANGXI UNIV
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