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Method for preparing in vitro fertilized dogs by using mature canine oocytes in vitro

An oocyte and in vitro maturation technology, applied in biochemical equipment and methods, germ cells, animal cells, etc., can solve the problems of low test success rate, limit the development and application of canine in vitro fertilization, increase the cost of experiments, etc., and achieve improvement Obtaining rate, reduction of animal production cost, effect of low cost

Active Publication Date: 2018-12-11
BEIJING SINOGENE BIOTECHNOLOGY CO LTD +1
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Problems solved by technology

Specifically, under normal circumstances, the accuracy rate of estrus identification is about 60%, and the test success rate is relatively low
Moreover, due to the need to use natural estrus female dogs as oocyte donors, the cost of experiments is greatly increased, which also greatly limits the development and application of canine in vitro fertilization (IVF) technology

Method used

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  • Method for preparing in vitro fertilized dogs by using mature canine oocytes in vitro
  • Method for preparing in vitro fertilized dogs by using mature canine oocytes in vitro
  • Method for preparing in vitro fertilized dogs by using mature canine oocytes in vitro

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Embodiment

[0041] 1. Collection and in vitro maturation of canine oocytes

[0042] Collect the sterilized ovaries of healthy bitches from the animal hospital, put them in a thermos cup filled with 37°C sterile saline supplemented with penicillin sodium and streptomycin sulfate, and transport them back to the laboratory as soon as possible within 3 hours. After receiving the test sample, use sterile scissors and forceps to carefully separate the ovarian tissue, clean up the connective tissue and fat around the ovary as much as possible, then transfer the ovary to a 50mL centrifuge tube, and use a sterile physiological Rinse with salt water three times. Put the cleaned ovarian tissue into a culture dish containing HM placed on a constant temperature platform at 37°C, cut the ovary with a sterile blade, and release oocytes as much as possible. Then select COCs under a stereomicroscope, and the selection criteria are: more than 2 layers of cumulus cells are wrapped, the diameter of the oocy...

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Abstract

The invention relates to a method for preparing in vitro fertilized dogs by using mature canine oocytes in vitro. The method comprises the steps of collecting the canine oocytes, and culturing the collected canine oocytes by using a maturation culture solution for performing in vitro maturation, wherein the maturation culture solution is a TCM199 culture solution in which 10-100mu g / ml of progesterone is added.

Description

technical field [0001] The invention relates to a method for preparing in vitro fertilized dogs by using in vitro matured canine oocytes, in particular to cultivating oocytes in vitro to obtain in vitro matured canine oocytes using maturation medium, and then co-cultured with capacitated sperm to obtain in vitro fertilization Eggs are then transplanted into recipient female dogs to obtain in vitro fertilized dogs. Background technique [0002] The reproductive physiology of dogs is relatively special. Unlike mammals such as cattle and sheep, dogs excrete immature oocytes in the GV stage during ovulation. Matures in the fallopian tube, therefore, it is difficult to obtain oocytes matured in vivo. At present, many mammals, including humans, can obtain embryos and offspring in vitro through certain channels, but dogs are different from other mammals. The rate of in vitro maturation (IVM) of canine oocytes has not been high, and the average MII (mature eggs) The rate is 10% to...

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Application Information

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IPC IPC(8): C12N5/075A61D19/04A01K67/027
CPCA01K67/027A61D19/04C12N5/0609A01K2227/10C12N2501/31C12N2500/30C12N2500/32
Inventor 郑敏米继东赵建平
Owner BEIJING SINOGENE BIOTECHNOLOGY CO LTD
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