Porcine Cumulus Oocyte Complex Isolation: A Technique to Isolate Cumulus Oocyte Complex from Porcine Ovarian Follicle for In Vitro Fertilization
Published: April 30, 2023
Abstract
Source: Gorczyca, G. et al., Proteolytically Degraded Alginate Hydrogels and Hydrophobic Microbioreactors for Porcine Oocyte Encapsulation. J. Vis. Exp. (2020).
This video describes the technique to isolate the cumulus-oocyte complex or COC from porcine ovaries. The isolated intact COC can be used to carry out the in vitro fertilization – the assisted reproductive technique for developing fertility treatments.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Isolation of porcine cumulus-oocyte complexes To collect material for the isolation of ovarian follicles, excise porcine ovaries from prepubertal gilts (approximately 6-7 months of age, weighing 70 to 80 kg) at a local slaughterhouse. Choose approximately 20 pig ovaries from 10 animals for cumulus-oocyte complexes (COCs) isolation in ea…
Declarações
The authors have nothing to disclose.
Materials
| General | |||
| Antibiotic Antimycotic (100x) 100ml | Thermo Fisher | 15240062 | 2.5% final concentration for Handling Medium. 1% in PBS (step 1.2) |
| DMEM/F12 (500ml) | Sigma-Aldrich | D8062 | Handling Medium |
| FCS (100 ml) | Thermo Fisher | 16140063 | 10% final concentration for Handling Medium. (step: 1.5) |
| PBS (1x, pH 7.4) 500ml | Thermo Fisher | 10010023 | |
| Maturation Medium | |||
| hCG (1 VIAL of 10 000 U) | Sigma-Aldrich | CG10 | |
| PMSG | BioVendor | RP1782721000 | |
| Specific instruments, tools | |||
| 30 mm Pteri dish | TPP | 93040 | |
| 60 mm IVF Petri dish | Falcon | 353653 | |
| Incubator | Panasonic | MCO-170AIC-PE | Other brands can be used |
| Sterile petri dish (10 cm) | NEST Biotechnology | 704002 | |
| Thermos | Quechua | 5602589 | Other brands can be used |
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Porcine Cumulus Oocyte Complex Isolation: A Technique to Isolate Cumulus Oocyte Complex from Porcine Ovarian Follicle for In Vitro Fertilization. J. Vis. Exp. (Pending Publication), e20842, doi: (2023).