Protocols for in vitro maturation of porcine oocytes usually involve collecting ovaries from an abattoir, transporting them to a laboratory in a phosphate-buffered saline solution at a fixed temperature and finally puncturing them. The oocytes retrieved are then sorted and matured. This study aimed to confirm that the in vitro maturation rate could be improved by puncturing the follicles directly on site (i.e., on the slaughter line) and then transporting their contents to the laboratory in a suitable medium at 37°C. To assess oocyte quality after maturation, we compared effects of abattoir puncture to those of laboratory puncture on the nuclear and cytoplasmic maturation rates of porcine oocytes after in vitro maturation. Oocytes were stained with Hoechst 33342 for the nuclear stage and FITC-PNA to assess cortical granule (GC) distribution and observed by confocal microscopy. The nuclear stages identified were the germinal vesicle, immature oocyte and mature oocyte. The oocytes were classified into three types according to the GC distribution pattern: immature, incomplete or fully mature. Follicular puncture performed at the abattoir and in the laboratory showed similar rates of nuclear maturation (90.7 ± 4.1% and 84.7 ± 6.2% respectively). However, the abattoir puncture showed a better cytoplasmic maturation rate than the laboratory puncture (45.3 ± 12.6% vs 19.0 ± 14.6%, P < 0.01). In addition, the abattoir puncture yielded higher-quality structures before maturation. It was also easier to maintain oocytes at 37°C than it was to maintain ovaries at room temperature.