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After centrifugation at 500g for 15 min (27, 70), semen samples from each group ( n = 3) were collected with the number of frozen-thawed spermatozoa adjusted to a final concentration of 1 10 6 sperms/ml, added to the above fertilization droplet containing MII oocytes (repeated in triplicate, 205 oocytes for the NC group, 210 oocytes for the GSH group, 220 oocytes for the SeNPs group and 217 oocytes for the GSH + SeNPs group, repeated in triplicate) and cultured in a CO 2 incubator (38.5C, 5%CO 2 and 5%O 2 ) for 18 h