Serum-free bovine oocyte in-vitro maturation culture solution and oocyte culture method
A technology for in vitro maturation culture and oocyte, which is applied in the field of bovine oocyte in vitro maturation medium and oocyte culture, and can solve the problem of lack of relevant reports on the research of serum-free culture system, and achieves the improvement of the early embryo development rate and the effect of maturity
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0025] Serum-free bovine oocyte maturation medium in vitro, including Hepes-free M199 basal medium, 6mg / mL fatty acid-free BSA, 0.075IU / ml HMG, 1μg / ml 17β-estradiol, 60ng / ml EGF at 0.57mM, L-cysteine at 0.57mM, bFGF at 40ng / ml, Glutamax (100x) at 1.6μL / ml, folic acid at 50μM, cholic acid at 2μg / mL and CXCL12 at 50ng / mL.
Embodiment 2
[0030] The mature culture of embodiment 2 bovine oocyte
[0031] The bovine ovaries were collected from designated slaughterhouses in Xi'an, Shaanxi Province. The ovaries were placed in a thermos flask containing 100 IU / ml penicillin and 100 μg / ml streptomycin in normal saline at 20-25°C, and transported back to the laboratory within 5 hours. After the ovary was transported back, use sterilized scissors to cut off the connective tissue, fat and attached fallopian tubes on the surface of the ovary, wash it three times in sterile normal saline, and use a 10mL syringe equipped with a 21G needle to extract the oocytes in the 2-8mm follicles on the surface of the ovary The cells were placed in a 6cm dish, and the cumulus-oocyte complexes (cumulus-oocyte complexes, COCs) were collected under a solid microscope. Wash three times in PBS after collection. COCs with normal oocyte morphology were selected for in vitro maturation culture.
[0032] Treatment group: the selected COCs were...
Embodiment 3
[0040] Embodiment 3 Preparation of bovine somatic cell cloned embryo
[0041] (1) Culture of bovine fetal fibroblasts
[0042] Take a tube of bovine fetal fibroblasts from passages 2 to 5 of Holstein cows (collected from the cattle farm of Yangling Keyuan Cloning Co., Ltd.) from liquid nitrogen and thaw at 39°C, add 0.8mL of DMEM / F12 cell culture medium, Centrifuge, discard the supernatant, add cell culture medium to resuspend, take 3mL of cell suspension and inoculate it in a 6cm-diameter petri dish, place in CO 2 Cultured at 38.5°C in an incubator.
[0043] When the bovine fetal fibroblasts reach 80% confluence, discard the culture medium and replace with Ca-free 2+ , Mg 2+ Rinse the cells with PBS, add trypsin and EDTA mixed digestion solution, and digest the cells. Observe the cells under an inverted microscope. When most of the cells retract, become round, and the intercellular space expands, stop digestion with DMEM / F12 cell culture medium containing 10% fetal bovine...
PUM
Property | Measurement | Unit |
---|---|---|
diameter | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com