L‐proline as a novel additive to cryopreservation media improved post‐thaw quality of human spermatozoon via reducing oxidative stress

In view of the detrimental effects of sperm cryopreservation, a routine technique in assisted reproductive technique (ART) laboratories, on spermatozoa, researchers herein examined if L-proline supplementation in cryopreservation medium could aid in improving sperm quality after cryopreservation.

• Normozoospermic men provided a total of 30 semen samples.

• The semen samples were cryopreserved using cryopreservation media supplemented with different concentrations of L-proline (0, 1, 2 and 4 mmol/L).

• Analysis of sperm parameters and chromatin integrity (aniline blue (AB), toluidine blue (TB), sperm chromatin dispersion test (SCD) and chromomycin A3 (CMA3)), reactive oxygen species (ROS), and total antioxidant capacity (TAC) and malondialdehyde (MDA) levels after thawing revealed significantly improved progressive motility and viability in correlation with supplementing sperm cryopreservation with a total of 4 mmol/L L-proline.

• Significantly diminished MDA and ROS levels were observed in samples cryopreserved using 4 mmol/L L-proline supplemented cryopreservation media.

• Also, it significantly increased TAC level. Also, samples cryopreserved using 4 mmol/L L-proline supplemented cryopreservation media showed significantly less chromatin damages (AB, TB and CMA3).

• Overall results support that the usage of L-proline supplemented cryopreservation media could aid in improving sperm quality after cryopreservation.