Pentoxifylline activates sperm motility in testicular sperm from patient with primary ciliary dyskinesia resulting in twin live birth through IVF-ICSI

Abstract

OBJECTIVE: Report live birth of healthy twins following IVF-ICSI with testicular sperm activated by pentoxifylline from a man with primary ciliary dyskinesia. DESIGN: Case report. MATERIALS AND METHODS: A 24-year old azoospermic male with primary ciliary dyskinesia (Kartagener's syndrome) and his 22-year old wife underwent IVF-ICSI with surgical retrieval of sperm. Testicular tissue was washed, placed in 1mL G-Sperm media(Vitrolife)+ 10% HSA(Vitrolife), minced, and incubated for 24hrs at 37°C with 6%CO2 resulting in <1million sperm/mL with 0% motility. Sperm were then incubated with 3.5mM pentoxifylline(Sigma) for 15min at 37°C with 6%CO2 resulting in slow, non-progressive motility. Of 16 oocytes retrieved, 8 were treated with hyaluronidase (80 U/mL)(Vitrolife) and denuded in G-Oocyte media(Vitrolife)+5%HSA at 37°C. ICSI dishes were prepared using 3-10μL drops of sperm-containing media+10μL drop of PVP(Vitrolife)+10μL drop of G-Oocyte media+5%HSA onto a Falcon1006 culture dish covered by Ovoil(Vitrolife). ICSI was performed on 8 oocytes which were washed and incubated in 0.5mL G1.3 media(Vitrolife)+5%HSA overlaid with 0.5mL Ovoil at 37°C with 6%CO2[sup] ; [/sup]8 oocytes were inseminated with donor sperm. Fertilization check was performed 15hrs post-ICSI. All 2PN embryos were cultured in 20μL G1.3 media+5%HSA overlaid with Ovoil for 48hrs. On Day 3 all embryos were transferred to 20uL G2.3 media(Vitrolife)+5%HSA overlaid with Ovoil and cultured to Day 5. All embryos were cultured at a density of 3-5embryos/drop in a 37°C incubator with 6%CO2. Embryo transfer was performed on Day 5 with a Wallace catheter(Irvine) containing 15μL of Embryo Glue(Vitrolife). RESULTS: Five of 8 oocytes injected with testicular sperm fertilized, 3 cleaved, and 2 hatching blastocysts were transferred on Day 5. The patient gave birth to healthy twins (males) at 36 weeks gestation. CONCLUSION: Pentoxifylline successfully activates testicular sperm from a male with primary ciliary dyskinesia resulting in twin live birth using IVF-ICSI.