A point of confusion for embryologists in the identification of viable spermatozoa by the eosin-nigrosin test / 대한생식의학회지

A viable spermatozoon is a prerequisite for fertilization in intracytoplasmic sperm injection (ICSI). Thus, it is crucial to select viable but immotile spermatozoa on the day of ICSI. We report conflicting results in the identification of viable but immotile spermatozoa between the eosin-nigrosin staining and the laser test, which resulted in confusion for embryologists during assisted reproductive technology (ART). Three patients’ semen samples that showed no motile spermatozoa are described in this report. To identify viable spermatozoa, we used both the eosin-nigrosin test and the laser test for each sample, and repeated the semen analysis twice in each patient. Viable but immotile spermatozoa selected by the laser test were used for ICSI. Viable spermatozoa were detected by both the eosin-nigrosin and laser tests in two patients (case 1, 95.00% vs. 24.21% and 92.68% vs. 22.22%; case 2, 41.18% vs. 23.48% and 39.81% vs. 22.52%), indicating consistent results between the two methods. In the third patient, the eosin-nigrosin test yielded viability rates of 20.75% and 19.14%, while the result of the laser test was 0%. Thus, testicular aspiration was performed to collect viable sperm from this patient. Normal fertilization was achieved after the injection of viable but immotile spermatozoa selected from these patients by the laser test, resulting in the birth of two healthy babies. Our study documents a case where the eosin-nigrosin test showed a limitation in identifying viable but immotile spermatozoa for ART, while the laser test may overcome this limitation. Larger samples may be required to corroborate the clinical value of the laser test.

A successful pregnancy using completely immotile but viable frozen-thawed spermatozoa selected by laser / 대한생식의학회지

The aim of this study was to report a successful pregnancy using completely immotile frozen-thawed spermatozoa selected by laser. A single laser shot was used to detect the presence of viable immotile spermatozoa in fresh and frozen-thawed testicular spermatozoa. The viability rate was 55.8% after the laser detection, and cryopreservation was carried out immediately. The thawing test was performed on the day of oocyte pick-up, and no motile sperm were observed after extending the culture for another 4 hours, while a survival rate of 39.8% was detected using the laser. In all, five mature oocytes were injected, resulting in four cases of normal fertilization (80%) on day 1. Further, two high-quality day 3 embryos were transferred, which resulted in a singleton pregnancy. Our study demonstrates that completely immotile spermatozoa are worth cryopreserving for further intracytoplasmic sperm injection, which provides a new insight into male fertility preservation in cases of completely immotile spermatozoa.

Influence of the insemination method on the outcomes of elective blastocyst culture / 대한생식의학회지

OBJECTIVE: The aim of this study was to explore the effects of the insemination method on the outcomes of elective blastocyst culture. METHODS: We retrospectively analyzed the outcomes of elective blastocyst culture performed between January 2011 and December 2014. RESULTS: There were 2,003 cycles of conventional in vitro fertilization (IVF) and 336 cycles of intracytoplasmic sperm injection (ICSI), including 25,652 and 4,164 embryos that underwent sequential blastocyst culture, respectively. No significant differences were found in the female patients' age, basal follicle-stimulating hormone level, basal luteinizing hormone level, body mass index, number of oocytes, maturity rate, fertilization rate, or good-quality embryo rate. However, the blastocyst formation rate and embryo utilization rate were significantly higher in the conventional IVF group than in the ICSI group (54.70% vs. 50.94% and 51.09% vs. 47.65%, respectively, p0.05). No cycles were canceled due to the formation of no usable blastocysts. CONCLUSION: Although the fertilization method had no effect on clinical outcomes, the blastocyst formation rate and embryo utilization rate in the ICSI group were significantly lower than those observed in the conventional IVF group. Therefore, more care should be taken when choosing to perform blastocyst culture in ICSI patients.

Effect of blastocyst quality on the strategy of single blastocyst transfer in frozen-thawed cycles / 中华妇产科杂志

Objective To investigate the effect of blastocyst quality on the strategy of single blastocyst transfer in frozen-thawed cycles. Methods A retrospective analysis was performed in Reproductive Medicine Center, Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region on clinical data of single frozen-thawed blastocyst transfer cycles from January 2008 to December 2013. All cycles were divided into four groups (AA, AB/BA, BB, BC/CB) according to the blastocyst score, then the clinical outcomes were compared between groups. And on this basis, the clinical outcomes were further explored when the group of outcomes with single blastocyst transfer wasn′t ideal, which would diverted to transfer two blastocyst. Results In single frozen blastocyst transfer cycles, the clinical pregnancy rate of each group with the blastocyst scored AA, AB/BA, BB, BC/CB were 61.4%(470/765), 51.2%(330/645), 40.5%(407/1 005), 22.9%(60/262), live births rate in each group were 52.2%(399/765), 41.2%(266/645), 30.4%(306/1 005), 13.7%(36/262), and the abortion rate were 13.6%(64/470), 16.7%(55/330), 21.4%(87/407), 35.0%(21/60), separately. This showed that the clinical pregnancy rate and live births rate decreased significantly with the decline of blastocyst quality (P40%) and live births rate (>30%) could be obtained, however, the clinical pregnancy rate of 22.9% and live births rate of 13.7%could only be acquired when blastocyst scored BC/CB only transferred one embryo, which significant lower than those of each group scored ≥BB grade (P0.05). Conclusions Of single blastocyst transfer in frozen-thawed cycles, the clinical pregnancy rate and liver births rate showed significant upward trend, but the abortion rate showed significant downward trend, with the decline of blastocyst quality. When the blastocyst scored ≥BB grade, the single blastocyst transfer could be considered to be performed.

Effects of artificial shrinkage of blastocoeles before vitrification on pregnancy outcome / 中华妇产科杂志

Objective To investigate the effects on pregnancy outcome and neonate by artificial shrinkage by microsucting the fluid of expanded blastocysts before vitrification using glass micropipette (GMP).Methods From Jan.2006 to Dec.2009, 342 vitrified-thawed blastocyst cycles from patients that performed in vitro fertilization-embryo transfer (IVF-ET) or intracyteplasmic sperm injection ( ICSI ) were enrolled in this study in Reproductive Medicine center, Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region.Three hundred and fourteen cycles of expanded blastocysts were artificially shranked by microsucting blastocoelic fluid with a micro-needle before vitrification as artificial shrinkage group, in the mean time, 28 cycles without artificial shrinkage were chosed as control group.The survival rate, implantation rate, clinical pregnancy rate and transfer canceled rate were compared between artificial shrinkage group and control group.Among pregnant women, the miscarriage rate, live birth rate, congenital birth defect rate, neonatal weight and gestational age were compared with those of fresh embryo transfers in 520 cycles.Results The blastocyst survival rate, implantation rate and clinical pregnancy rate were 95.3%(403/423), 38.0% ( 153/403), 44.6% (140/314) in artificial shrinkage group and 64.3 % (27/42),7.4% (2/27), 7.1% (2/28) in control group, respectively, which reached statistical difference (P＜0.05).The transfer canceled rate was 0 in artificial shrinkage group and 25.0% (7/28) in control group, which also reached statistical difference ( P ＜ 0.05 ).Among pregnant patients, the miscarriage rate of 18.2% (10/55), live birth rate of 80.0% (44/55), gestational age of (38.2 ± 1.3) weeks, congenital birth defect rate of 2.1% (1/47), birth weight of newborns of (2989 ±640) gram in artificial shrinkage group were not significantly different with 17.5% (91/520), 74.0% (385/520), (37.9 ±2.3) weeks,1.7% (8/479) and (2856±640) gramin fresh embryo transfer group (P＞0.05).Conclusion Artificial shrinkage by microsucting blastocoelic fluid with a micro-needle before vitrification significantly improved the vitrification effects of expanded blastocyst and no distinct increasing rate of neonates congenital anomality were observed.