ABSTRACT
This study was performed to evaluate the efficiency of simplified EM grid vitrification, skipping the step of removing the cryoprotectant (5.5M EG + 1.0M sucrose) droplet on the grid after loading oocytes, compared to conventional cryopreservation protocols for mouse mature oocytes. Firstly, the recovery, survival, fertilization and hatching rates of simplified EM grid vitrification were compared with those of the slow freezing method using 1.5M DMSO. Then, conventional EM grid vitrification was compared with simplified EM grid vitrification. Simplified EM grid vitrification showed higher survival, fertilization and hatching rates than those of the slow freezing method (85.6% vs. 63.2%; 51.0% vs. 22.3%; 38.7% vs. 12.5%, p < 0.01, respectively). Moreover, simplified EM grid vitrification showed higher recovery, survival and fertilization rates than those of conventional EM grid vitrification (100% vs. 95.0%, p=0.024; 90.0% vs. 78.9%, p=0.033; 56.7% vs. 38.7%, p=0.021, respectively). Hatching rate tended to be higher for simplified EM grid vitrification compared to conventional EM grid vitrification (41.1% vs. 24.1%). In conclusion, simplified EM grid vitrification is a convenient and efficient method for cryopreservation of mouse mature oocytes, compared to conventional EM grid vitrification and slow freezing methods.
Subject(s)
Pregnancy , Mice , Male , Female , Animals , Oocytes/cytology , Mice, Inbred DBA , Mice, Inbred C57BL , Fertilization in Vitro , Cryopreservation/instrumentation , Cell SurvivalABSTRACT
OBJECTIVE: This study was carried out to compare the clinical outcomes of intracytoplasmic sperm injection (ICSI) in patients with obstructive azoospermia according to sperm retrieval site and technique; microsurgical epididymal sperm aspiration (MESA), percutaneous epididymal sperm aspiration (PESA), testicular sperm extraction by open biopsy (TESE). METHODS: The outcomes of ICSI and IVF-ET were evaluated and compared among 3 groups. Seventy three men suffering from infertility due to obstructive azoospermia had 107 ICSI cycles using MESA (21 cycles in 15 patients), PESA (26 cycles in 17 patients) and TESE (60 cycles in 41 patients). RESULTS: In the clinical outcomes in patients undergoing ICSI with epididymal or testicular sperm, there were no significant differences in fertilization rate (66.1% vs. 60.5%), cleavage rate (94.9% vs. 97.6%), cumulative embryo score (CES) (51.3 vs. 58.8), implantation rate (7.9% vs. 6.1), and clinical pregnancy rate per ET (30.4% (14/46) vs. 25.4% (15/59)) between both groups. Also, in the clinical outcomes in ICSI patients using MESA, PESA, TESE, there were no significant differences in fertilization rate (61.8%, 69.4%, 60.5%), cleavage rate (92.1%, 97.3%, 97.6%), CES (38.1, 52.0, 58.8), implantation rate (9.5%, 6.6%, 6.1%), and clinical pregnancy rate per ET (35% (7/20), 26.9% (7/26), 25.4% (15/59)) among 3 groups. CONCLUSION: When compared with MESA or TESE, PESA, the clinical outcomes were similar in ICSI patients with obstructive azoospermia whatever the origin or the technique of sperm retrieval. However, we considered PESA is more time-saving and cost effective for ICSI in patients with obstructive azoospermia.
Subject(s)
Humans , Male , Azoospermia , Biopsy , Embryonic Structures , Fertilization , Infertility , Pregnancy Rate , Sperm Injections, Intracytoplasmic , Sperm Retrieval , SpermatozoaABSTRACT
OBJECTIVE: This study was carried out to compare the clinical outcomes of intracytoplasmic sperm injection (ICSI) in patients with obstructive azoospermia according to sperm retrieval site and technique; microsurgical epididymal sperm aspiration (MESA), percutaneous epididymal sperm aspiration (PESA), testicular sperm extraction by open biopsy (TESE). METHODS: The outcomes of ICSI and IVF-ET were evaluated and compared among 3 groups. Seventy three men suffering from infertility due to obstructive azoospermia had 107 ICSI cycles using MESA (21 cycles in 15 patients), PESA (26 cycles in 17 patients) and TESE (60 cycles in 41 patients). RESULTS: In the clinical outcomes in patients undergoing ICSI with epididymal or testicular sperm, there were no significant differences in fertilization rate (66.1% vs. 60.5%), cleavage rate (94.9% vs. 97.6%), cumulative embryo score (CES) (51.3 vs. 58.8), implantation rate (7.9% vs. 6.1), and clinical pregnancy rate per ET (30.4% (14/46) vs. 25.4% (15/59)) between both groups. Also, in the clinical outcomes in ICSI patients using MESA, PESA, TESE, there were no significant differences in fertilization rate (61.8%, 69.4%, 60.5%), cleavage rate (92.1%, 97.3%, 97.6%), CES (38.1, 52.0, 58.8), implantation rate (9.5%, 6.6%, 6.1%), and clinical pregnancy rate per ET (35% (7/20), 26.9% (7/26), 25.4% (15/59)) among 3 groups. CONCLUSION: When compared with MESA or TESE, PESA, the clinical outcomes were similar in ICSI patients with obstructive azoospermia whatever the origin or the technique of sperm retrieval. However, we considered PESA is more time-saving and cost effective for ICSI in patients with obstructive azoospermia.
Subject(s)
Humans , Male , Azoospermia , Biopsy , Embryonic Structures , Fertilization , Infertility , Pregnancy Rate , Sperm Injections, Intracytoplasmic , Sperm Retrieval , SpermatozoaABSTRACT
OBJECTIVE: To establish the optimal cryopreservation method in mouse oocytes. METHODS: Firstly, mouse immature oocytes were exposed to various cryoprotectants, and then cryoprotectant with the best outcome was selected. Secondly, mouse immature oocytes were cryopreserved by either slow freezing and ultra-rapid thawing or vitrification. Finally, in mouse mature oocytes, the five different protocols were compared in their fertilization and hatching rates. RESULTS: 1) 1.5M 1,2-propanediol (PROH) and 1.5M PROH+0.1M sucrose had a higher rate of survival (73.1%, 81.9%) and in vitro maturation (28.2%, 30.1%). 2) Vitrification using 5.5M ethylene glycol (EG) showed significantly higher rate of survival and in vitro maturation, when compared with slow freezing and ultra-rapid thawing using 1.5M PROH+0.1M sucrose (65.9% vs 50.0%, 40.0% vs 28.2%, respectively). 3) In mouse mature oocytes, vitrification using 5.5M EG showed significantly higher survival rate, however, slow freezing and ultra-rapid thawing using 1.5M DMSO was superior to vitrification in view of fertilization rate. CONCLUSIONS: Vitrification showed better outcomes in mouse immature oocytes, but slow freezing and ultra-rapid thawing using 1.5M DMSO may be beneficial in mature oocytes.