Method for improving the embryo transplantation pregnancy rate for cattle and sheep

A technology of embryo transfer and sheep embryo, which is applied in the field of embryo transfer, can solve the problems of restricting the application of embryo transfer technology and high production cost, and achieve the effect of low application cost, simple operation and remarkable effect

Inactive Publication Date: 2008-02-06
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the high genetic quality and production cost of transplanted embryos, the death of early embryos brings huge economic losses, which directly limits the application of embryo transfer technology in production
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Method used

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  • Method for improving the embryo transplantation pregnancy rate for cattle and sheep
  • Method for improving the embryo transplantation pregnancy rate for cattle and sheep
  • Method for improving the embryo transplantation pregnancy rate for cattle and sheep

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1. Adding Bovine Recombinant IFN-tau to the Transplant Fluid to Improve the Pregnancy Rate of Cow Transplantation

[0024] In this case, by means of genetic engineering, by cloning the bovine IFN-tau gene, constructing an expression vector, and further transforming host cells to produce IFN-tau protein, from which IFN-tau is isolated and purified, and added to the transplantation fluid to improve embryo transfer. pregnancy rate.

[0025] (1) Cloning and prokaryotic expression of bovine IFN-tau gene

[0026] 1. Embryo Lysis

[0027] (1) Take out the thin tube containing the bovine male embryo from the liquid nitrogen, put it in a water bath at 32°C for 10 seconds to thaw, and take out the thin tube;

[0028] (2) Cut off the sealing end of the thin tube, push the cotton plug with a metal push rod, so that the contents in the tube together with the embryos flow into the watch glass containing 1×PBS (or Holding solution) solution;

[0029] (3) quickly recover emb...

Embodiment 2

[0056] Example 2 Embryo co-implantation

[0057] In this case, normal embryos and embryos with developmental defects were co-implanted to increase the level of INF-tau. The developmental defect embryo refers to using in vitro matured bovine ovum to induce parthenogenetic development with chemical reagents, and cryopreservation after developing into a blastocyst. During embryo transfer, parthenogenetic embryos and normal embryos are injected into the uterus at the same time in a ratio of 1:1. IFN-tau secreted by parthenogenetic embryos can promote normal embryonic development and implantation.

[0058] (1) Bovine oocyte collection and in vitro maturation culture

[0059] The bovine ovaries obtained from the slaughterhouse were placed in normal saline at 25°C and brought back to the laboratory. Use a 10mL syringe with a 19G needle to extract the oocytes in 2-6mm follicles, select the complete and dense COCs (cumulus granulosa cells and oocyte complexes) under a solid microsco...

Embodiment 3

[0069] Example 3 Co-transplantation of embryonic tissue fragments

[0070] The method in this case is the co-transplantation of normal embryos with living tissue fragments secreting IFN-tau.

[0071] In this case, the inner cell mass of the in vivo fertilized blastocyst that was no longer used after sex identification was used, and only the trophoblast cell mass was retained. During embryo transfer, normal embryos and trophoblast cell blocks (containing about 60 cells) were co-transplanted into the uterus of recipient cows, and the IFN-tau secreted by trophoblast cells would promote normal embryo development and implantation. Using the above method to transfer 40 embryos, 28 of them were implanted, and the pregnancy rate reached 70%, which was 15% higher than that of the control group.

[0072] social and economic benefits

[0073] As can be seen from the above examples, the method of the present invention can significantly improve the pregnancy rate after embryo transfer of...

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Abstract

The invention provides a method for increasing embryo transplantation pregnancy rate of cattle and sheep, which increases embryo transplantation pregnancy rate through improving IFN-tau level inside maternal uterus. The invention also provides a plurality of feasible methods for improving IFN-tau level, which comprise adding IFN-tau into transplantation liquid during embryo transplantation and transplanting parthenogenesis embryo or living tissue fragments together with normal embryo. Through the method provided by the invention, embryo transplantation pregnancy rate of cattle and sheep can be substantially increased by 15 percent to 20 percent as compared with the control group; meanwhile, the invention has a simple operation, low application cost, outstanding effect without any harmful influence on animal, thereby being suitable for extensive application in cattle and sheep embryo transplantation industry and embryo engineering and having huge market potential.

Description

technical field [0001] The invention relates to embryo transfer technology, in particular to a method for improving the embryo transfer pregnancy rate of cattle and sheep. Background technique [0002] Embryo transfer technology can make full use of the genetic resources of fine breeding stock and rapidly expand the proportion of fine breeds in the herd. At the same time, this technology is also an effective means of treating infertility in mammals. However, the efficiency of livestock embryo transfer technology still has not reached the ideal level. For example, the conception rate of fresh embryos in dairy cows is only 55%-60%, the conception rate of thawed embryos after freezing is only 45%-50%, and the conception rate of in vitro fertilized embryos is 30%. %, while the conception rate of somatic cell nuclear transfer cloned embryos is only 10%-20%. The efficiency of embryo transfer in sheep and goats is basically the same. However, the pregnancy rate of natural mating...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61D19/04
Inventor 曾申明
Owner CHINA AGRI UNIV
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