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Firstly, to determine an adequate concentration of caffeine for separation, frozen\u2013thawed sperm were incubated with different concentrations of caffeine (0, 1, 2.5, 5, and 10\u2009mM) in a MIGLIS device. To determine the appropriate incubation time for separating sperm in the MIGLIS device, frozen\u2013thawed sperm were incubated with 2.5\u2009mM caffeine for 5, 10, 15, or 20\u2009min. To evaluate the fertilization and embryo development of oocytes fertilized with frozen\u2013thawed sperm separated into two regions (outer and inner) in the MIGLIS device, the separated sperm from the three boars was used to fertilize in\u00a0vitro\u2010matured oocytes and cultured in\u00a0vitro for 7\u2009days. Sperm quality parameters of sperm collected from the inner tube after incubation with 2.5\u2009mM caffeine were superior to sperm incubated without caffeine. Moreover, sperm collected from the inner tube after incubation for 10\u2009min had a higher progressive motility. The rate of blastocyst produced from spermatozoa collected from the inner tube after incubation with 2.5\u2009mM caffeine for 10\u2009min significantly increased compared to that produced from spermatozoa from the outer tube, regardless of the boar. In conclusion, sperm sorting using the MIGLIS device may be useful for separating high\u2010quality sperm after incubation with 2.5\u2009mM caffeine for 10\u2009min to improve blastocyst formation.<\/jats:p>","DOI":"10.1111\/rda.14648","type":"journal-article","created":{"date-parts":[[2024,6,15]],"date-time":"2024-06-15T07:49:00Z","timestamp":1718437740000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Quality and fertilizing ability of frozen\u2013thawed porcine sperm separated using a migration sedimentation method"],"prefix":"10.1111","volume":"59","author":[{"given":"Suong Thi","family":"Nguyen","sequence":"first","affiliation":[{"name":"Joint Graduate School of Veterinary Sciences Yamaguchi University  Yamaguchi Japan"},{"name":"Bio\u2010Innovation Research Center Tokushima University  Tokushima 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