Case presentation
The couple presented with 4 years of primary infertility. The 33-year-old woman had been assessed by a gynaecologist, and no contributing female factors were detected. After physical examination, the 34-year-old man was normal in the male reproductive system,and no significant respiratory symptoms were observed. His history provided no further information of clinical problems,and lymphocyte karyotype was 46, XY. Analysis of microdeletions of azoospermia factor genes on Y chromosome was normal.However,after repeated semen analyses by light microscope, the man was subjected to complete immotility and totally abnormal tail, namely MMAF with short, cureled, bent, coiled, absent flflagellum(Figure1A). According to the World Health Organization guidelines,Semen analyses showed sperm densities of 11-21 million/mL, volumes of 2.6–3.6 mL, and sperm vitality of 39% with normal pH.The sperm flagellar displayed curled 64%, bent20%, short 7%, absent 3%, coiled 2%, irregular 4%(Table1). The couple had not previously attempted IVF.
A long protocol for ovulation induction was administrated by the daily administration of recombinant FSH (200 IU/day) following pituitary desensitisation with GnRH agonist. and cumulus-oocyte complexes were retrieved transvaginally 36 h later under ultrasound guidance after administration of 10,000 IU of human chorionic gonadotrophin (HCG). Oocytes were identified and maintained in culture under 6% CO2 in humidified air at 37°C .
After the oocyte retrieval procedure,cumulus-oocyte complexes were digested by using hyaluronidase (10 IU/ml). 22 of the 24 oocytes were confirmed as being at the metaphase II stage of meiosis . 10 mature oocytes were injected with ejaculated sperm and other 12 of them were vitrifed in the event of future ICSI attempts with testicular serpmatozoa. In the fresh cycle,6 of the 10 microinjected oocytes were fertilized with ejaculated sperm(Table1).On day 3 , two good quality embryos in culture were transferred. the other four non-good quality embryos did not develop to blastocyst after six days . Pregnancy was not achieved after 14 days of embryos’ transfer.
Seven months later, the couple returned to the clinic in order to try again with their frozen oocytes and fresh testicular spermatozoa.Testicular spermatozoa were obtained by needleaspiration. Compared to ejaculated sperm,there was nothing changed about the testicular spermatozoa morphology,which showed that all were 100% immotile and curled or bent back on itself. Testicular sperm with comparatively normal morphology were injected to 12 warmed oocytes, which survied sucessfully after thawing. Nine two-pronuclei and one-pronuclei oocytes were obtained at 16 h post-ICSI.cleavage was observed in eight oocytes.on day 3,two 8-cell embryos with < 20% fragmentation were transferred.two of the remaining 6 embryos developed to blastocyst after D6,and a grade of 5BB blastocyst was frozened(Table1).Twelve days after transfer, the βHCG concentration was 80 mIU/dl. But a gestational sac with fetal heartbeat was not observed 3 weeks later.
Fortunately,offspring were obtained in the last attempt after transfer the frozen blastocyst of 5BB. A normal female baby was delivered,with a birth weight of 3050g and a length of 53cm.