Clinical and neonatal outcomes of using a modified micro cryotube for cryopreservation of small numbers of spermatozoa for TESA-ICSI cycles.

Cryopreservation of a small number of human spermatozoa is still a major challenge for embryologists. The aim of this study was to evaluate the clinical pregnancy and neonatal outcomes of intracytoplasmic sperm injection (ICSI) using a modified micro cryotube as freezing carrier for freezing small numbers of human spermatozoa collected by testicular sperm aspiration (TESA). We conducted a retrospective study to analyses the ICSI outcomes of using frozen-thawed few testicular spermatozoa in males with obstructive azoospermia (OA) from June 2017 to June 2021. Of 155 ICSI treatment cycles, 79 cycles were allocated to frozen sperm group and a modified micro cryotube was used for freezing testicular sperm, 76 cycles were allocated as fresh sperm group. No significant differences were observed in fertilization rate, good quality embryo rate, and blastocyst rate between the frozen sperm group and fresh sperm group (P > 0.05). Similarly, in the fresh embryo transfer cycles plus the first frozen-thawed embryo transfer cycles, the total clinical pregnancy rate (54.43% vs 57.89%), implantation rate (46.08% vs 49.47%), miscarriage rate (13.95% vs 13.64%) and live birth rate (45.57% vs 48.68%) were not statistically different between the frozen and fresh sperm groups (P > 0.05). In addition, there was no statistical differences in the mean gestational age (38.33weeks ± 1.74 vs 37.89weeks ± 1.87), preterm delivery rate (5.56% vs 10.81%), mean birth weight at delivery (3026.50 g ± 577.64 vs 2977.56 g ± 528.93), and low birth weight (12.50% vs 19.51%) between the two groups (P > 0.05 in all cases). Modified micro cryotube for cryopreservation of rare testicula rretrieved spermatozoa did not negatively affect the pregnancy and neonatal outcomes in TESA-ICSI cycles. The presented method may be a useful alternative for cryopreservation of small numbers of human spermatozoa in clinical setting.

Laser-assisted selection of immotile spermatozoa has no effect on obstetric and neonatal outcomes of TESA-ICSI pregnancies.

BACKGROUND: Azoospermic patients have benefited from both epididymal and testicular spermatozoa intracytoplasmic sperm injection (ICSI) treatment and lasers have been used to identify viable, immotile spermatozoa before the procedure. There are limited studies on the safety of laser-assisted selection of immotile spermatozoa. The aim of this study was to investigate the impact of laser-assisted selection of immotile spermatozoa on the obstetric and neonatal outcomes after ICSI. METHODS: A retrospective comparative study was conducted on outcomes of ICSI cycles with testicular spermatozoa from June 2014 to June 2018. Of 132 cycles, 33 were allocated to the test group and oocytes were injected with immotile spermatozoa selected by laser, 99 cycles were allocated as control group. RESULTS: Compared with the control group, no significant differences were found in the pregnancy, implantation, miscarriage and live birth rates in the test group in either fresh or frozen transfer cycles. The cumulative live birth rate in the test group was 69.70%, which was slightly higher than in the control group (60.61%), but this was not statistically different. There were no differences in the average gestational age, premature birth rate, neonatal birth weight, and the malformation rate between the test and control groups (P > 0.05). In addition, the obstetric outcome between the two groups were not different (P > 0.05). CONCLUSIONS: No negative effect on perinatal and neonatal outcomes was seen by using laser-assisted selection of immotile spermatozoa for TESA-ICSI. This study endorses the use of laser-assisted selection of viable spermatozoa for ICSI cycles.

Does longer storage of blastocysts with equal grades in a cryopreserved state affect the perinatal outcomes?

AIM: Although mammalian embryos could be preserved in liquid nitrogen for thousands of years in theoretical models, the viability of cryopreserved blastocyst with varying grades remains to be speculated. In this study, we aimed to determine whether the longer storage time of blastocysts with equal grades could negatively affect the perinatal outcomes. MATERIALS AND METHODS: Single vitrified-warmed blastocyst was divided into four grades (AA, AB/BA, BB, BC/CB) according to the blastocyst score when freezing, and each grade of blastocyst was categorized into four storage duration categories: 28 days-1 year, 1-3 years, 3-5 years, and ≥5 years. Then the perinatal outcomes with different storage time were analyzed. RESULTS: Our results revealed that for blastocysts with the same grade, the length of storage time had no statistical effect on blastocyst survival rate, clinical pregnancy/implantation rate, live birth rate, and abortion rate. In addition, more advanced developmental blastocyst could obtain better pregnancy outcomes regardless of the cryopreservation length. Similar neonatal outcomes were obtained over time. CONCLUSIONS: Cryopreservation time could not negatively affect the perinatal outcomes of blastocysts with equal grades. Efficient blastocyst cryopreservation technology by vitrification can help older women obtain high-quality embryos at a young age.

Choosing the optimal blastocyst by morphology score versus developmental rate in frozen-thawed embryo transfer cycles.

The aim of this study was to provide guidance for better management in the selection of blastocyst to warm in frozen-thawed embryo transfer (FET) cycles. A retrospective cohort follow-up study was conducted that included single autologous frozen blastocyst transfer cycles performed in our Reproductive Medicine Unit from January 2009 to December 2016. The live birth rate (LBR), clinical pregnancy rate (cPR) were increased as blastocyst morphology scores increased, but the miscarriage rate decreased in all groups. In the high-score groups, there were no differences in LBR between D5 and D6, while in the low-score groups, LBR was significantly higher in D5 compared to the D6. With respect to neonatal outcome, there were no differences in all the groups. After binary logistic regression analysis, it was seen that patients' age, thawed cycles, pre-frozen morphology score and developmental rate were independently associated with LBR. These results suggest that for high-scoring blastocyst, the pre-frozen morphological score should be given priority while for low-scoring blastocysts, the developmental rate should be given priority when thawing in FET cycles.

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

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.

Cumulus oophorus complexes favor physiologic selection of spermatozoa for intracytoplasmic sperm injection.

OBJECTIVE: To investigate the effectiveness of cumulus oophorus complexes (COCs) in the physiologic selection of spermatozoa for intracytoplasmic sperm injection (ICSI). DESIGN: A prospective sibling oocytes study. SETTING: Center of reproductive medicine. PATIENT(S): Couples undergoing ICSI during 2016, females aged ≤38 years, and at least six metaphase II (MII) oocytes retrieved. Sixty patients were included in the study. Of 857 MII oocytes, 429 were allocated to the study group and were injected with the sperm selected via COCs; 428 MII oocytes were allocated as controls (C) and fertilized by conventional ICSI. INTERVENTION(S): In the study group, ICSI was performed with spermatozoa that traversed the COCs in vitro. MAIN OUTCOMES MEASURE(S): Blastocyst/top blastocyst formation rate, fertilization rate, and oocyte utilization rate. RESULT(S): Oocytes injected with COC-selected spermatozoa had a significantly higher fertilization rate than the conventional ICSI group (85.31% vs. 74.77%). There were no statistically differences in cleavage and top embryo rate on day 3 between the COC-ICSI and C-ICSI groups. However, with day 5 or 6 embryos, compared with conventional ICSI, COC-ICSI significantly improved blastocyst formation rate (64.90% vs. 53.50%), blastocyst formation rate at day 5 (46.52% vs. 38.85%), top blastocyst rate (38.72% vs. 24.20%), and the usable blastocysts formation rate (62.12% vs. 46.82%). The oocyte utilization rate was improved greatly in the COC-ICSI group compared with the C-ICSI group (51.98% vs. 34.35%). Furthermore, the fertilization rate, top embryo rate on day 3, usable blastocyst rate, top blastocyst rate, and day 5 usable blastocysts rate were similar between the conventional IVF and COC-ICSI groups. Single-blastocyst transfer was performed in 82 cycles, including 44 fresh cycles and 38 frozen-thawed cycles. The cumulative embryo implantation rate in the COC-ICSI group was 64.29%, slightly higher than in the C-ICSI group (53.85%), but there was no statistical difference. CONCLUSION(S): The use of COCs to select spermatozoa for ICSI appears to be effective and led to a statistically significant improvement in blastocyst development and quality.

A new insight into male fertility preservation for patients with completely immotile spermatozoa.

BACKGROUND: Sperm cryopreservation is the most effective method to preserve male fertility but this is normally used for motile spermatozoa. Thus, only motile spermatozoa are used for cryopreservation in most reproductive medicine centers worldwide. The immotile spermatozoa from some problematic patients are usually discarded, resulting in a missed opportunity of sterility cryopreservation for future assisted reproductive treatments. Many studies have shown that successful fertilization can be obtained after selection of viable sperm from the completely immotile spermatozoa before ICSI. Whether the completely immotile spermatozoa are worth of freezing has not been realized The aim of this study is to explore the clinical value of cryopreservation of immotile spermatozoa. METHODS: Completely immotile spermatozoa were collected and frozen, and subsequently viable but immotile frozen-thawed spermatozoa were selected by laser plus for ICSI. Main outcomes included spermatozoa survival index, fertilization rate and good quality embryo rate. RESULTS: After identification by laser, the fresh samples of spermatozoa presented with a mean survival rate of 54.86% and 26.05%, and this was reduced to 44.13% and 18.13% in frozen-thawed spermatozoa samples, which showed a frozen-thawed spermatozoa survival index of 0.80 and 0.70 in the testicular and ejaculate sperm, respectively. There were no statistically differences in fertilization rate (80% vs80.51%, 75.00% vs 81.48%), cleavage rate (95.45% vs 98.95%, 100.00% vs 95.45%) and good quality embryo rate (40.48% vs 52.13%, 33.33%vs38.10%) between the frozen-thawed immotile spermatozoa group and the routine fresh immotile spermatozoa ICSI group in both testicular and ejaculate sperm, respectively. CONCLUSIONS: The results of the study show that completely immotile spermatozoa can be frozen in order to preserve male fertility as long as viable spermatozoa are present. This procedure provides a further possibility for fertility preservation for patients with completely immotile spermatozoa.

Influence of the insemination method on the outcomes of elective blastocyst culture.

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, p<0.05). The implantation/pregnancy rate (IVF, 50.93%; ICSI, 55.10%), miscarriage rate (IVF, 12.57%; ICSI, 16.29%), and live birth rate (IVF, 42.12%; ICSI, 44.08%) were similar (p>0.05). 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.

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

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.

Blastocoele re-expansion time in vitrified-warmed cycles is a strong predictor of clinical pregnancy outcome.

AIM: To assess the predictive value of blastocoele re-expansion time in clinical pregnancy outcome in vitrified-warmed cycles. METHODS: Data on 468 single vitrified-thawed blastocyst transfer cycles (in patients aged <38 years) carried out from January 2012 through December 2012, at the Reproductive Medicine Center, Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region, were analyzed. Vitrified-warmed blastocysts were divided into three groups according to blastocoele re-expansion time: group A, <1 h; group B, 1-2 h; and group C, >2 h, and the clinical pregnancy outcomes (i.e. live birth rate, miscarriage rate and occurrence of singleton pregnancies) compared between the groups. RESULTS: Significant differences were observed in the implantation/clinical pregnancy rate between groups A, B and C (70.10%, 51.76% and 28.74%, respectively, P < 0.01). There was a significant linear decline in this rate with increasing blastocyst re-expansion time. The rate of miscarriage also tended to increase with increasing blastocyst re-expansion time, but the difference was not statistically significant (P > 0.05). Of the pregnant patients, no significant difference was observed in the rates of monozygotic twins and ectopic pregnancy between the three groups. For the newborns, similar live birth, low-birthweight and premature delivery rates were observed between the groups. CONCLUSIONS: Timing of blastocoele re-expansion in vitrified-warmed cycles is a strong predictor of clinical pregnancy outcome. The faster the re-expansion of the blastocoele, the higher the developmental potential of the blastocysts.

[Successful pregnancy following laser-assisted selection of viable but immotile spermatozoa for intracytoplasmic sperm injection: A report of 2 cases].

OBJECTIVE: To investigate the feasibility and clinical application value of selecting viable spermatozoa by noncontact diode laser. METHODS: We obtained immotile spermatozoa from 2 infertile men with obstructive azoospermia or severe asthenospermia and selected viable spermatozoa using a single laser shot at the sperm tail. Those that responded to the laser shot by a curling reaction of the tail were regarded as presumably viable and used for intracytoplasmic sperm injection (ICSI). RESULTS: The mean fertilization rate was 88.89% after ICSI with the laser-selected viable spermatozoa. Both of the embryo transfers resulted in a single pregnancy. CONCLUSION: Noncontact diode laser is a useful alternative for the assessment of sperm viability, which may help to achieve successful pregnancy.

[Effects of IVF versus ICSI on the outcomes of elective blastocyst culture].

OBJECTIVE: To explore the effects of different fertilization methods on the outcomes of elective blastocyst culture. METHODS: We retrospectively analyzed the outcomes of elective blastocyst culture for 1 153 cycles of IVF and 205 cycles of ICSI performed between january 2009 and December 2012. RESULTS: A total number of 14 748 embryos in the IVF group and 2 655 embryos in the ICSI group underwent sequential blastocyst culture, with 7 871 blastocysts formed in the former and 1 210 in the latter. No cycles were canceled for no blastocyst formation in either of the two groups. The rates of quality embryos, blastocyst formation and embryo utilization were significantly higher in the IVF than in the ICSI group (64.77 vs 58.72%, 53.37 vs 45.57%, and 60.06 vs 52.17%, all P < 0.05), but the rates of implantation, clinical pregnancy and abortion showed no significant differences between the two groups (48.94 vs 51.43%, 49.03 vs 52.02%, and 11.69% vs 15.56, all P > 0.05). CONCLUSION: With the same inclusion criteria of selective blastocyst culture, IVF has a lower risk of cycle cancellation due to no blastocyst formation and therefore may effect higher rates of blastocyst formation and embryo utilization than ICSI. Our study suggested that appropriate inclusion criteria of selective blastocyst culture should be laid down according to different fertilization methods.

[Predictive value of sperm morphology according to WHO Laboratory Manual for the Examination and Processing of Human Semen (5th Ed) on the outcomes of IVF-ET].

OBJECTIVE: To determine the predictive value of sperm morphology based on the criteria of the 5th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen (WHO5) on the outcomes and neonatal status following IVF-ET. METHODS: According to the strict criteria of WHO5, we obtained semen samples from 789 subjects and allocated them to a normal group (morphologically normal sperm > or = 4%, 754 cycles) and a teratozoospermia group (morphologically normal sperm < 4%, 35 cycles). We made comparisons between the two groups in the rates of normal fertilization, cleavage, quality embryo, implantation, clinical pregnancy and miscarriage as well as the status of the neonates. RESULTS: No significant differences were observed in the couples' age, mean number of oocytes, and mean stature and body mass index of the women between the two groups (P > 0.05). The teratozoospermia group showed slightly lower rates of fertilization, cleavage, quality embryo, embryo cryopreservation, implantation and pregnancy, but a higher rate of miscarriage than the normal group (P > 0.05). Apart from 141 on-going pregnancies (140 in the normal and 1 in the teratozoospermia group), 228 healthy infants were born following 789 transfer cycles, 213 (141 singletons and 36 twins) in the former and 15 (9 singletons and 3 twins) in the latter group. Congenital defects were found in none of the neonates, and there were no significant differences in the gestation period, premature birth rate and low body weight between the two groups (P > 0.05). CONCLUSION: Sperm morphology according to the criteria of WHO5 has but a limited value in predicting the outcomes and neonatal status following IVF-ET.

Comparable clinical outcomes and live births after single vitrified-warmed and fresh blastocyst transfer.

Selective single-blastocyst transfer (SBT) in fresh cycles has been effective in reducing multiple pregnancies. However, we do not know whether this successful strategy of fresh transfer cycles is suitable for cryopreserved cycles. The present study was undertaken to evaluate the feasibility and value of SBT in vitrified-warmed cycles. Clinical pregnancy rate (CPR) was similar with vitrified and fresh SBT (46.61% versus 52.15% respectively). Of the pregnant patients, monozygotic twin, miscarriage and ectopic pregnancy rates were similar with vitrified and fresh SBT. For the newborns, no significant difference was observed in live birth, low birthweight, premature delivery and birth defects rates between vitrified and fresh SBT. With respect to the quality of transferred blastocysts (from BB to AA), a similar CPR and miscarriage rate was obtained for both vitrified and fresh SBT when a similar blastocyst cohort graded ≥ 3BB was transferred. The data show that vitrified SBT is an effective means of reducing multiple pregnancy and that comparable clinical outcomes and live births are achieved if single blastocysts graded ≥ 3BB are transferred for both vitrified and fresh SBT. These data should encourage clinics to evaluate their embryo transfer policy and adopt vitrified SBT as everyday practice. Selective single-blastocyst transfer in fresh cycles has been an effective method to reduce the multiple pregnancies. However, due to a lack of adequate studies, we do not know whether this successful strategy in fresh transfer cycles is suitable in cryopreserved cycles. The present study was undertaken to explore the feasibility and value of single-blastocyst transfer in vitrified-warmed cycles. We found that single-blastocyst transfer in vitrified-warmed cycles is an effective means of reducing multiple pregnancy, and comparable clinical outcomes and live births were achieved if single blastocysts graded ≥ 3BB were transferred for both vitrified-warmed and fresh blastocyst transfer. These data should encourage clinics to evaluate their embryo transfer policy and adopt single-blastocyst transfer in cryopreserved cycles as their everyday practice.

[Effects of artificial shrinkage of blastocoeles before vitrification on pregnancy outcome].

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 intracytoplasmic 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) gram in 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.

[Influence of sperm morphology on the outcomes and neonatal status in IVF-ET].

OBJECTIVE: To determine the influence of sperm morphology on the outcomes and status of the neonates in in vitro fertilization and embryo transfer (IVF-ET). METHODS: Strictly based on the WHO criteria, we divided semen samples into a moderately abnormal group (sperm of normal morphology 5% - 10%), a mildly abnormal group (10% < sperm of normal morphology < 15%) and a normal group (sperm of normal morphology > or = 15%) , and compared the rates of fertilization, cleavage, quality embryos, implantation, clinical pregnancy and live births among the three groups. RESULTS: There were not significant differences in the patients' age among the three groups (P > 0.05). The fertilization rate was markedly lower in the moderately abnormal than in the mildly abnormal group (63.70% vs 73.74%, P < 0.05), but not significantly different from the normal group (63.70% vs 68.05%, P > 0.05). The rate of quality embryos of the normal group was the highest, significantly higher than that of the moderately abnormal group (44.83% vs 35.75%, P < 0.05), but no statistically significant differences were observed in the rates of cleavage, implantation and clinical pregnancy among the three groups (P > 0.05). A total of 125 babies were born from the 280 ET cycles, including 73 singletons and 26 twins, of whom none showed any congenital birth defects. No statistically significant differences were found in the rates of abortion, ectopic pregnancy and premature delivery, nor in the mean gestational period and average body weight of the neonates among the three groups (P > 0.05). CONCLUSION: Moderately abnormal sperm morphology did not affect the fertility rate of IVF, but significantly decreased the quality of embryos; mildly abnormal sperm morphology had no obvious influence on the rates of fertilization, cleavage, quality embryos, implantation, clinical pregnancy and live births; while normal sperm morphology played a limited role in predicting the outcomes and status of the neonates in IVF-ET.