Application Of Micro-amount Testicular Spermatozoa Cryopreservation In Assisted Reproduction

Objective:To evaluate the efficiency of micro-amount spermatozoa cryopreservation and to compare the clinical pregnancy outcomes in intracytoplasmic sperm injection(ICSI)using fresh versus cryopreserved spermatozoa collected by testicular sperm aspiration(TESA)in patients with azoospermia.Methods:Ultra-low temperature freezing technology is widely used in human assisted reproduction,including egg cells,sperm,and embryo freezing.Normal male sperm freezing technology is very mature and widely used in all major human sperm banks.However,the sperm obtained after the operation of TESA in spermatozoa patients has a low sperm density and poor survival rate,and the use of conventional sperm freezing methods is not ideal.This study explored the microsperm freezing technology,combined with ICSI in severe male factors infertility patients,for clinical treatment.To review and analyze 273 cycles of clinical data from the patients with azoospermia who received ICSI treatment in the Reproductive Hospital of Shandong University from January 2015 to December 2016.The 79 cycles using frozen-thaw testicular sperms for ICSI were classified into frozen TESA sperm group.The 194 cycles using fresh TESA-derived sperm was classified as fresh TESA sperm group.The rates of 2PN fertilization,good-quality embryos,blastocyst formation,implantation and clinical pregnancy outcomes were obtained from clinical records and were compared between the two groups.Results:1.The female age,male age,the body mass index,infertility period,the level of FSH,E2 on hCG trigger day,total dose of Gn,duration of stimulation,endometrial thickness and other ICSI characteristics of the two groups were similar(p>0.05).2.There was no significant difference on the rates of 2PN fertilization,good-quality embryos,good-quality blastocyst between the Frozen TESA sperm group(73.4%?54.5%?60.1%)and fresh TESA sperm group(71.4%?55.3%?60.9%)(P>0.05).3.Compared the rates of blastocyst formation and good-quality blastocyst from poor quality embryos in the two groups.The rates of blastocyst formation and good-quality blastocyst in the frozen TESA sperm group(56.7%?29.0%)were slightly higher than fresh TESA sperm group(50.4%?27.2%),but the difference was not statistically significant(p>0.05).4.There was no significant difference on the rates of implantation,clinical pregnancy,spontaneous abortion,live delivery between the frozen TESA sperm group(42.7%?55.1%?14.8%?45.7%)and fresh TESA sperm group(47.3%?61.7%?1 3.6%?51.1%)(P>0.05).Conclusion:ICSI using cryopreserved-thawed micro-amount sperm collected by TESA can obtain very good clinical pregnancy outcomes.Using the cryopreserved-thawed sperm for ICSI can avoid damage to the testicles by repeated percutaneous testicular puncture,also reduce the economic and mental burden of patients.So it is a safe and effective method in the treatment of patients with azoospermia.But the effect of ICSI using cryopreserved-thawed sperm on the neonatal development and health requires more basic research and clinical outcome tracking to explore and verify.Significance:Microsperm freezing can reduce the physical injury of male patients after repeated surgery,and greatly preserves the fertility of patients,which has a very high clinical application value.At present,the domestic center of reproduction officially carried out this technology is not many,this study for the application of microsperm freezing in clinical,provided a relatively rich clinical data and experience,for the development of microsperm freezing technology provides a good idea.For the first time,straw was used as a refrigerated carrier,combined with a refrigerated protective agent to freeze microsperm of testis.There was no significant difference between the recovery rate of spermatozoa with that of normal spermatozoa.After a large number of preliminary experiments,spenn recovery rate is high,and the operation is simple,suitable for the actual work needs of the laboratory.