We investigated the feasibility of agarose-gel microcapsules to cryopreserve extremely small numbers of sperm for assisted reproductive technology. Semen samples were collected from 16 patients attending the center for reproductive medicine male infertility clinic at a university hospital. We used agarose microcapsules to cryopreserve extremely small numbers of sperm from 16 patients with male infertility (10 with sperm concentration >= 1 million/mL; 6 with sperm concentration <1 million/mL). Six spermatozoa were injected into agarose-gel microcapsules and cryopreserved in a liquid nitrogen tank for 7 days. The Crytop method was used for cryopreservation as a control. After thawing, spermatozoa were recovered. Sperm recovery rates, motility and viability, and recovery time were compared. The post-thawing recovery rate, motility rate, and viability rate were higher whereas the recovery time was shorter in samples preserved using the agarose-gel microcapsule method compared to samples preserved using the Cryotop method in both the group with sperm concentrations of 1 million/mL or above and the group with sperm concentrations of less than 1 million/mL. This study demonstrated that using the agarose-gel microcapsule method increased post-thawing sperm recovery rate, sperm motility rate, and sperm viability rate, and reduced sperm recovery time compared with the conventional Cryotop method when cryopreserving samples with low sperm count. Although requiring further study, the agarose-gel microcapsule method shows much promise as a new option for freezing sperm.
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