Method for improving blastocyst rate of in-vitro embryos of animals
A technology of embryonic development and animal body, applied in embryonic cells, animal cells, vertebrate cells, etc., to reduce production costs, improve early development ability, and improve production efficiency.
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Embodiment 1
[0022] Example 1 The effect of rbIFN-τ on the development of parthenogenetically activated embryos in bovine oocytes
[0023] 1. Collection and in vitro maturation of bovine oocytes
[0024] Bovine ovaries were collected from the slaughterhouse and placed in thermos flasks containing physiological saline at 27±2°C. After arriving at the laboratory, wash the ovaries with sterilized saline preheated at 37°C for 3 times, and then use a syringe to aspirate the cumulus-oocyte complexes (COCs) in the 2-8mm follicle above the ovary, and the complete cumulus cells will be collected. The layer or part of the dense cumulus cells was washed twice in the oocyte washing solution, and then washed three times in the maturation solution, and the washed oocytes were transferred into 100 μL oocyte maturation culture microdrops in vitro, and each microdrop was placed Insert 15~20 oocytes, and finally put them in 38.5℃, 5%CO 2 and cultured in a carbon dioxide incubator with saturated humidity f...
Embodiment 2
[0034] Example 2 The effect of rbIFN-τ on the development of bovine oocyte fertilized embryo in vitro
[0035] 1. In vitro fertilization of bovine oocytes
[0036] (1) Preparation of mature oocytes before in vitro fertilization
[0037] See item 1 in Example 1 for the collection and in vitro maturation of bovine oocytes. Transfer the oocytes matured in vitro for 24 hours into pre-balanced BO fertilization solution (38.5°C, 5% CO 2 (incubator) for 3 washes, and then transferred to 50 μL of fertilized fluid droplets in a pre-balanced 60 mm Petri dish and placed at 38.5 °C, 5% CO 2 15-20 mature oocytes per fertilized microdrop.
[0038] (2) Preparation of sperm
[0039] After thawing the frozen semen in a water bath at 37°C, take 0.25 μL of semen and carefully place it in a sterilized 10 mL centrifuge tube containing 7 mL of BO capacitation solution, then centrifuge (1800 rpm, 5 minutes) to wash twice, and then place under the microscope Adjust sperm concentration to 2×10 wi...
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