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Updated: Dec 25, 2025

Mouse Sperm Cryopreservation and Recovery using the I·Cryo Kit
Published on: December 12, 2011
[L-carnitine protects the motion parameters and mitochondrial function of human sperm in cryopreservation]
Long-Long Fu1,2, Lin-Yuan Zhang3, Qi An1,2
1Graduate School, Peking Union Medical College, Beijing 100730, China.
Objective:
To study the effects of L-carnitine (LC) on cryopreserved human sperm.
Methods:
Ten semen samples were collected from normal sperm donors, each divided into six groups, fresh ejaculate (FE), non-LC cryopreservation (non-LC), and cryopreservation with LC at 1 mmol/L (LC-1), 2.5 mmol/L (LC-2), 5 mmol/L (LC-3) and 10 mmol/L (LC-4), respectively. The optimal concentration of LC was identified based on the motility and motion parameters of the post-thaw sperm. The plasma membrane integrity (PMI) of the sperm was assessed by eosin-nigrosin staining, their mitochondrial membrane potential (MMP) monitored by JC-1 assay, and the level of sperm ROS measured by the fluorescent probe DCFH-DA, followed by analysis of the mechanisms of LC protecting sperm against cryopreservation injury.
Results:
Compared with the sperm in the FE group, the post-thaw sperm in the non-LC and LC groups showed significantly decreased progressive motility, average path velocity (VAP), straight line velocity (VSP) and curvilinear velocity (VCP) (P < 0.05). In comparison with the non-LC group, the LC-3 group exhibited a remarkably higher percentage of progressively motile sperm ([41.9 ± 4.6] vs [47.0 ± 4.3]%, P = 0.0261) and VAP ([34.9 ± 2.6] vs [38.9 ± 4.2] μm/s, P = 0.0152), indicating that the optimal concentration of LC was 5 mmol/L. Both PMI and MMP were significantly lower in the non-LC than in the FE group ([52.7 ± 5.7] vs [75.5 ± 5.4]%, P < 0.01 and [44.5 ± 3.5] vs [57.3 ± 4.4]%, P < 0.01), but higher in the LC groups ([70.1 ± 8.2]% and [50.3 ± 3.4]%) than in the non-LC group (P < 0.01 and P < 0.05). The level of sperm ROS, however, was markedly higher in the non-LC than in the FE group ([12.5 ± 3.9] vs [6.8 ± 2.4], P < 0.01) but lower in the LC groups ([8.4 ± 5.3]%) than in the non-LC group (P = 0.05).
Conclusions:
L-carnitine can improve the motility and motion parameters of cryopreserved human sperm by reducing sperm ROS, enhancing sperm mitochondrial membrane potential and protecting the sperm plasma membrane.
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