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).

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