One milliliter of each sample of liquefied semen was mixed with 2 mL of Ham’s F10 (Sigma, USA), supplemented with 20% human serum albumin (HSA) (Kedrion Biopharma, Italy),
and centrifuged at 300 g for 10 min. The supernatant was decanted, and 1 mL of Ham’s F10 was added. The tubes were gently positioned at a 45-degree angle for one
hour at 37 °C and 5% CO2 for swim-up separation of motile spermatozoa. This supernatant was carefully aspirated, and its volume was adjusted to 1 mL using Ham’s F10.
and centrifuged at 300 g for 10 min. The supernatant was decanted, and 1 mL of Ham’s F10 was added. The tubes were gently positioned at a 45-degree angle for one
hour at 37 °C and 5% CO2 for swim-up separation of motile spermatozoa. This supernatant was carefully aspirated, and its volume was adjusted to 1 mL using Ham’s F10.
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