Comparison of postthawing sperm motility recovery between cryopreserved with and without cryoprotective agent using 4 different cryopreservationmethods
Hilwah Nora Shofwal Widad Irwan Taufiqur Rachman
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Abstract
Frozen-thawed human spermatozoa are routinely used for many assisted reproduction program. However, cryopreserved spermatozoa was reported to yield lower pregnancy rates compared to fresh semen in both intra uterine inseminations and in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) due to the reduction of sperm motility and viability induced by cryopreservation procedures. This study was aimed to evaluate the influence of cryoprotective agent (CPA) and cryopreservation methods on human sperm motility. This was a quasi experimental study. Thirty seven normozoospermic semen samples collected in Permata Hati Infertility Clinics of Dr. Sardjito General Hospital, Yogyakarta were recruited. Four different cryopreservation methods were applied using and without CPA (TEST-yolk buffer). In simple two steps freezing, cryostraw were gradually frozen from 8 to -4°C. In simple graduated freezing, cryostraus were directly frozen at -4°C. In vapor phase freezing method, the samples in cryostraw were placed 1 cm above liquid nitrogen. In the last method, the samples were directly submerged into liquid nitrogen. Thawing was conducted by incubation at 37°C for 5 minutes. The sperm motility recovery after cryopreservation in the 4 different cryopreservation methods was evaluated and analyzed by analysis of variance (ANOVA). The fresh sperm motility before cryopreservation was 52.9 ± 4.50%. The recovery of motile sperms was 17.00 ± 7.83%, 20.96 ± 5.81%, 15.06 ± 8.55% and 15.68 ± 8.3%, when using CPA and 5.63 ± 4.63%, 5.47 ± 3.95%, 4.45 ± 4.46% and 6.08 ± 5.06% when without CPA following direct plunge to liquid nitrogen freezing, vapor liquid nitrogen freezing, simple graduated freezing and simple 2-steps freezing, respectively. Among methods using CPA, the vapor phase method resulted in highest sperm motility recovery. In methods without CPA, no significant difference of sperm motility recovery was observed among the 4 different cryopreservation methods. In conclusion, the use of CPA for cryopreservation improves sperm motility recovery.
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