Cultivation method of cross-bred wagy

A technology of bull and bovine embryos, applied in the preservation of human or animal bodies, medical science, embryonic cells, etc., to achieve the effect of improving meat quality standards and soft texture

Pending Publication Date: 2013-07-03
AIDE INSPECTION TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using the method of vitrification to freeze in vitro embryos has the advantages of simple operation, low cost, and high hatching rate of embryos after thawing, but the disadvantage is that the operator is required to have the skills to master this method

Method used

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  • Cultivation method of cross-bred wagy
  • Cultivation method of cross-bred wagy
  • Cultivation method of cross-bred wagy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] The acquisition and maturation culture of embodiment 1 oocyte

[0053] The oocytes obtained from the ovaries of Canadian Holstein cows collected from Canadian slaughterhouses were processed in the laboratory. The operating procedures are as follows: (a) The equipment for collecting and shipping ovaries must be cleaned in advance; (b) Prepare high-pressure sterile saline in advance, add 1% antibiotics before use, and keep warm in a water bath at 28°C until Use; (c) Collect cow ovaries with no obvious lesions and no feces contamination, and put the ovaries into a thermos bottle with pre-packed physiological saline; (d) The time from the collection of ovaries to their utilization shall not exceed 10 hours .

[0054] The procedure for extracting oocytes is as follows: (a) transfer the ovary to a wide-mouth thermos flask, pour in saline solution with 1% antibiotics at about 30°C until the ovary is submerged, and use it after cleaning the ovary; (b) 10mL syringe cover Put o...

Embodiment 2

[0061] Example 2 In vitro fertilization

[0062]The in vitro fertilization culture solution of the present invention (called 603 culture solution, its formula is shown in Table 3. The specific formula of 603 culture solution in the present embodiment is: add 50 mL of hydroxyethylpiperazine ethanesulfonic acid, heparin 0.6mL, fructose 1.2g, lactic acid 10mL, calf serum albumin 6g, phenol red 0.02g, penicillin 50IU, streptomycin 50IU, after mixing, add culture mother solution to 1000mL, adjust the pH value to 7.2) is conventional fertilization culture The improved version of TALP can change the oocyte cytoplasm to be more suitable for sperm penetration, and focus on the capacitation efficiency of sperm. Two hours before in vitro fertilization of in vitro embryos, take a four-well plate, and add 600 microliters of 603 culture medium to each well, and then put it in 39°C, 5% CO 2 incubator for pre-incubation. The frozen semen treatment in the cultivation method of the present in...

Embodiment 3

[0067] Embodiment 3 culture embryo in vitro

[0068] The configuration of the embryo culture solution of the present invention is added on the basis of the 601 culture mother solution (see Table 1 for the formula). In the present invention, the embryo culture solution is referred to as 604 culture solution for short, and the formula is shown in Table 5. In this example, the specific formula of the 604 culture solution is: 3g of glycine, 2g of alanine, 3g of arginine, 2g of glutamic acid, 2g of cystine, 5g of lysine and 5g of methionine were added to 800mL of culture mother solution , 5 g of leucine, 5 g of phenylalanine, 60 g of fetal bovine serum albumin, 0.3 g of inositol, 50 IU of penicillin, and 40 IU of streptomycin. Finally, it was sterile filtered and stored at 4°C for later use. Two hours before in vitro embryo culture, take a four-well plate and add 680 microliters of 604 culture solution to each well, then add 320 microliters of sterilized paraffin oil to the cultu...

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Abstract

The invention provides a cultivation method of cross-bred wagy. The method comprises the steps of culturing cross-bred wagy embryo in vitro and transplanting the cultured embryo into uterus of receptor cow to breed, wherein the embryo in-vitro culture method comprises the following steps: (1) taking an ox with 99.9% of Japanese wagy hereditary characters as a paternal line, and Holstein cow as a maternal line, respectively extracting semen of the paternal line and ovocyte of the maternal line; (2) culturing the ovocyte in ovocyte follicle fluid; (3) performing in-vitro fertilization on mature ovocyte in an in-vitro fertilization culture solution; (4) culturing the fertilized ovum in the embryo culture solution; and (5) vitrifying the in-vitro embryo. The in-vitro cultured embryo is transplanted into the uterus of receptor cow by a bilateral uterine lining embryo transplantation method. At last, the cross-bred first filial generation wagy new strain with characters of Japanese wagy and Holstein cow is cultivated, and has high table purpose value and good production performance.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for breeding hybrids and cattle. Background technique [0002] Japanese Wagyu is a long-term breed of beef cattle that Kobe, Japan has been isolated from the outside world for centuries. Japanese Wagyu is recognized as an excellent breed of beef cattle in the world. The quality of the beef produced by this breed is also unique. Its characteristics are: soft texture, tender and juicy, strong beef flavor, and the appearance of the beef shows strong marbling , distributed in the beef evenly and uniformly like snowflakes in a multi-star shape, fresh and delicious, rich in iron and unsaturated fatty acids, which can significantly improve energy and endurance, and delay the aging of the body. It can be called the best in beef products. The Japanese government has regarded Japanese Wagyu as a national treasure. In the 1990s, the Japanese government issued a ban on "exporting Wagy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073C12N5/075A01N1/02A61D19/04A01K67/027
Inventor 祝加贝
Owner AIDE INSPECTION TECH
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