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Lowline embryo freezing liquid, freezing method and unfreezing method

A freezing method and bovine embryo technology, which is applied in the field of bioengineering and embryo genetics, can solve the problems of survival rate and pregnancy rate impact, limit the development of bovine embryo transfer technology, etc., and achieve the effect of simple operation, strong operability and convenient operation

Inactive Publication Date: 2014-12-17
南宁培元基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using the above three cryoprotectants to freeze bovine embryos, the freezing effects of embryos are not the same, and the survival rate and pregnancy rate after thawing are affected to varying degrees, which has become a major problem that limits the further development of bovine embryo transfer technology.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] 1. Obtaining of Embryos

[0033] Use non-surgical methods to flush the donor cattle that have undergone superovulation treatment. First, fix the donor cattle in a safety frame, and inject 5% lidocaine hydrochloride into the epidural cavity of the sacral-tail junction of the donor cattle. (Reasonable injection according to the weight of the cow) After anesthesia, embryo punching (collecting 6-8 days old morula or early blastocyst from the uterus). Put the ovum collection cup filled with the egg recovery solution under a stereo microscope, observe carefully from low power to high power, and put the detected embryos into a φ35 mm petri dish filled with preservation solution.

[0034] The quality identification is carried out under a stereo microscope, and the identification is carried out in strict accordance with the embryo quality identification standards. The excellent and good grade embryos are frozen.

[0035] 2. Embryo pretreatment

[0036] After washing the embry...

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Abstract

The invention discloses a lowline embryo freezing liquid, a freezing method and an unfreezing method and belongs to the technical field of bioengineering and embryonic heredity. The lowline embryo freezing liqui comprises ethanediol, cane sugar and DMSO. The freezing method comprises the following steps: freezing an obtained embryo for pre-treating, putting the embryo in a tube and freezing the embryo. The unfreezing method comprises the following steps: retaining a straw in air for 5-10 seconds, putting the straw in water bath at the temperature of 35-38 DEG C for 10-15 seconds, detecting the embryo, transferring the embryo in WM1 liquid drops, transferring into another hole liquid drops containing the same solution, then respectively transferring to WM2 and liquid drop containing HM with the concentration being 800 microliter per hole and retaining for 5 minutes each time; and washing the embryo by a culture solution for three times, continuously observing the development condition of the embryo in the culture solution or transplanting the embryo to a receptor cow after being subjected to estrus synchronization. The freezing liquid, the freezing method and the unfreezing method have the good freezing effect, are strong in operability, convenient to operate and suitable for popularization and application.

Description

[0001] technical field [0002] The invention relates to the technical fields of bioengineering and embryo genetics, in particular to a freezing liquid, a freezing method and a thawing method for Loulai bovine embryos. Background technique [0003] Embryo cryopreservation technology is a key link in the stage of industrialization, application and development of embryo transfer technology. This technology can make embryo transfer not limited by time and region, simplify the cost of live animal transportation and introduction, and prevent the spread of epidemic diseases. This technology can also be used to establish embryo banks of fine breeds of domestic animals and endangered animals, effectively preserving the genetic resources of various fine breeds and endangered species. At present, in the study of cryopreservation of embryos, it is mainly to seek low-toxicity antifreeze protective agents, effective and reasonable, easy-to-operate methods, and the determinants of embryo...

Claims

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Application Information

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IPC IPC(8): A01N1/02
Inventor 蒋和生
Owner 南宁培元基因科技有限公司
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