Apparatus for collecting early-stage embryo or early-stage oocyte of pig
A technology of oocytes and embryos, applied in veterinary instruments, medical science, animal delivery, etc., can solve the problems of lower egg flushing rate, egg flushing failure, easy bleeding, etc., and achieve the effect of improving the success rate and operating freely
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Embodiment 1
[0030] Embodiment 1 Embryo Collection and Transplantation of Excellent Breeding Pigs
[0031]Using high-quality purebred sows as donors and hybrid sows that cannot be used as breeding recipients, the embryos of donor sows are transplanted to the recipients of hybrid sows, with the purpose of producing high-quality purebred pigs from hybrid sows. The operation steps are as follows:
[0032] 1. Superovulation and mating of donor sows
[0033] On the 15th-18th day of the estrous cycle of the donor sow, intramuscularly inject 1000 IU of the serum gonadotropin PMSG of the pregnant horse, and inject 800 IU of the human chorionic gonadotropin HCG 72 hours later for superovulation treatment. After estrus, the boars of this breed are used for breeding.
[0034] 2. Simultaneous estrus of recipient sows
[0035] On the 15th-18th day of the estrous cycle of the recipient sow, while the superovulation treatment of the donor sow, PMSG 800 IU was injected intramuscularly, and HCG 600 IU...
Embodiment 2
[0055] Example 2 Preparation of transgenic pigs with human serum albumin
[0056] 1. Preparation of human serum albumin gene
[0057] Cut the constructed human serum albumin gene plasmid into linearity, and dilute it with TE diluent to a 5 μg / ml gene solution for later use.
[0058] 2. Collection of Embryos
[0059] Using the same method as in Example 1, the sows were subjected to superovulation treatment and mated after estrus; 24 hours after the last mating, surgery, anesthesia, stabilization, and abdominal opening were carried out, and the process was the same as in Example 1. The operator extends the index finger and middle finger into the abdominal cavity through the incision, touches the uterus or uterine horn at the front and rear positions at the junction with the pelvic cavity, grasps it with two fingers, pulls it to the surface of the wound, and leads out the fallopian tube along one side of the uterine horn. Find the ovary at the end of the fallopian tube. Firs...
Embodiment 3
[0071] Example 3 Construction of EGFP gene cloned pigs
[0072] 1. Preparation of donor cells
[0073] Take out the cryopreservation tube of transgenic porcine fetal fibroblasts from liquid nitrogen, put it in a 37°C water bath and shake carefully to make it thaw quickly; remove the cryopreservation solution by centrifugation, add 2mL culture medium with a pipette, and gently suspend cell. Adjust the cell density to 10 6 cells / mL, and cultured in DMEM containing 0.5% fetal bovine serum for 48-72 h with serum starvation. Afterwards, they were digested with trypsin and collected, and placed in the modified NCSU-23 phosphate culture medium for storage at 38.5°C for 20-120 minutes before being used for nuclear donors.
[0074] 2. Preparation of oocytes
[0075]Sows are not mated after estrus. Within 12 hours after the end of estrus, anesthesia and Baoding are used to make an incision, and the oviduct and ovary are drawn out of the incision; one end of the oviduct is inserted...
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