The influence of clinical and laboratory factors on the formation of monopronucleated zygotes after intracytoplasmic sperm injection (ICSI).

SummaryThe aim of the present study was to determine whether clinical or laboratory factors can influence the development of single pronucleated zygotes (1PN) and two polar bodies (PB) after ICSI. In total, 341 ICSI cycles performed at FertiClinic-Villa Margherita from January 2012 to December 2014 were enrolled in the study. Group A included 240 cycles with no 1PN-2PB while group B included 101 cycles with one or more 1PN-2PB. Age, stimulation protocol, infertility factor, amount of gonadotropin administered, duration of therapy, peak estradiol levels, number of follicles at maturation triggering, oocytes retrieved and mature oocytes, time between retrieval and injection and sperm characteristics were compared between groups. In opposition to previous results showing no relationship between 1PN occurrence and clinical or laboratory variables, we observed that 1PN-2PB zygote formation seems to be associated with a lower female age, higher level of E2 and higher number of follicles on day of oocyte maturation triggering, higher number of astenozoospermic male patients, more oocytes retrieved at pick-up, more mature oocytes (MII) and longer time to injection.

Evaluation of the Efficiency of Two Different Freezing Media and Two Different Protocols to Preserve Human Spermatozoa from Cryoinjury.

It is universally recognized that cryopreservation impairs sperm quality. In order to improve postthawing sperm survival and motility, media of different composition and different protocols have been proposed. However, no clear evidence is available to understand which are the most efficient protocol and medium for sperm cryopreservation. The present study evaluates the efficiency of two different cryopreservation protocols and two common freezing media (FM) containing different cryoprotectants (CPs), TEST Yolk Buffer (TYB) and Sperm Freeze (SF), to preserve human sperm quality. Our data suggest that TYB is better than SF both in terms of postthaw viability and in terms of progressive motility, while the direct addition of FM to the sperm sample resulted in the most efficient protocol in terms of postthaw viability but not in terms of progressive motility.

Morphological assessment on day 4 and its prognostic power in selecting viable embryos for transfer.

The aim of this study was to describe a system for embryo morphology scoring at the morula stage and to determine the efficiency of this model in selecting viable embryos for transfer. In total, 519 embryos from 122 patients undergoing intracytoplasmic sperm injection (ICSI) were scored retrospectively on day 4 according to the grading system proposed in this article. Two separate quality scores were assigned to each embryo in relation to the grade of compaction and fragmentation and their developmental fate was then observed on days 5 and 6. Secondly, the prediction value of this scoring system was compared with the prediction value of the traditional scoring system adopted on day 3. Morulas classified as grade A showed a significant higher blastocyst formation rate (87.2%) compared with grades B, C and D (63.8, 41.3 and 15.0%, respectively), (P < 0.001). Furthermore, the ability to form top quality blastocysts was significantly higher for grade A morulas with respect to grades B, and C and D (37.8% vs. 22.4% vs. 11.1%), (P < 0.001). Finally, the morula scoring system showed more prediction power with respect to the embryo scoring a value of 1 [Akaike information criterion (AIC) index 16.4 vs. 635.3 and Bayesian information criterion (BIC) index -68.8 vs. -30.0 for morulas and embryos respectively]. In conclusion, results demonstrated that the presented scoring system allows for the evaluation of eligible embryos for transfer as a significant correlation between the grade of morula, blastulation rate and blastocyst quality was observed. Furthermore, the morula scoring system was shown to be the best predictive model when compared with the traditional scoring system performed on day 3.

Cetrorelix protocol versus gonadotropin-releasing hormone analog suppression long protocol for superovulation in intracytoplasmic sperm injection patients older than 40.

OBJECTIVE: To determine which protocols work better between cetrorelix and long protocols in older patients in a randomized controlled study. DESIGN: A controlled randomized study in a single private IVF center. SETTING: Infertile women referred to a private IVF center. PATIENT(S): Five hundred sixty-four women 40 years or older undergoing IVF. INTERVENTION(S): At their first IVF cycle, the women were randomized into two study groups using a computer-generated number sequence: 281 cases were treated with the cetrorelix protocol, and 283 patients were treated with a long protocol for controlled ovarian hyperstimulation. MAIN OUTCOME MEASURE(S): Days of stimulation, E(2) on the day of hCG administration, amount of FSH administered, number of oocytes yielded, number of embryos obtained, pregnancy rate, and implantation rate. RESULT(S): Patients treated with the long protocol showed a significantly higher number of oocytes retrieved and a higher pregnancy rate for both the cycle and transfer with respect to the cetrorelix protocol patients. The other parameter evaluated did not show any statistically significant differences. CONCLUSION(S): Our study showed that the long protocol performed better in older women than the cetrorelix protocol and that the GnRH antagonist may be detrimental in older women.

Natural-cycle in vitro fertilization in poor responder patients: a survey of 500 consecutive cycles.

OBJECTIVE: To determine the role of the natural cycle for in vitro fertilization (IVF) in poor responder patients. DESIGN: Retrospective survey. SETTING: Private center for assisted reproduction. PATIENT(S): 294 women who were poor responders in a previous IVF cycle. INTERVENTION(S): Analysis of 500 consecutive natural cycles IVF. MAIN OUTCOME MEASURE(S): Number of cycles with oocytes, pregnancy rate per cycle, per transfer, and implantation rate. RESULT(S): Oocytes were found in 391 cases (78.1%), and cleaving embryos suitable for transfer were obtained in 285 cycles (57.0%). Pregnancy was observed in 49 cases, with a pregnancy rate of 9.8% per cycle, 17.1% per transfer, and 16.7% per patient. The patients were subdivided arbitrarily by the women's age into three groups. Patients 35 years old or younger showed a pregnancy rate of 18.1% per cycle, 29.2% per transfer, and 31.7% per patient. Women aged between 36 and 39 years showed a pregnancy rate of 11.7% per cycle, 20.6% per transfer, and 20.3% per patient. Women 40 years old or older showed a pregnancy rate of 5.8% per cycle, 10.5% per transfer, and 9.7% per patient. No differences were found for any of the evaluated parameters, independent of which cycle was the first, the second, third, fourth, or fifth, or further consecutive cycle. CONCLUSION(S): In poor responder patients, natural-cycle IVF is an effective treatment, especially in younger women.

Use of a medium buffered with N-hydroxyethylpiperazine-N-ethanesulfonate (HEPES) in intracytoplasmic sperm injection procedures is detrimental to the outcome of in vitro fertilization.

OBJECTIVE: This study was conducted to determine whether N-hydroxyethylpiperazine-N-ethanesulfonate (HEPES)-buffered medium used for the microinjection of sperm into oocytes may be detrimental for the embryo. DESIGN: Controlled randomized study. SETTING: Private IVF center. PATIENT(S): Women (n = 708) undergoing ICSI. INTERVENTION(S): The women were randomized into two study groups: 2,204 oocytes from 357 women were treated using a medium buffered with bicarbonate without HEPES during the ICSI procedure, and 2,168 oocytes from 351 women were treated using a medium buffered with HEPES during the ICSI procedure. MAIN OUTCOME MEASURE(S): Fertilization rate, degeneration rate, triploid rate, cleavage rate, embryo quality, pregnancy rate, implantation rate, and abortion rate. RESULT(S): Oocytes treated with a HEPES-buffered medium showed a statistically significant higher rate of triploid and degenerated oocytes after fertilization with ICSI compared with oocytes treated with a medium without HEPES. The embryos obtained from oocytes microinjected with a HEPES-buffered medium showed a statistically significant higher rate of highly fragmented embryos compared with the controls. Pregnancy rate and implantation rate were statistically significantly lower in the patient group with oocytes treated with the HEPES-buffered medium. The other parameters evaluated did not show any statistically significant differences. CONCLUSION(S): Our study showed that the use of media buffered with HEPES, during the microinjection of sperm into the oocytes, is detrimental for IVF outcome and should be avoided.

A controlled trial of natural cycle versus microdose gonadotropin-releasing hormone analog flare cycles in poor responders undergoing in vitro fertilization.

OBJECTIVE: To determine the efficacy of natural-cycle IVF compared with controlled ovarian hyperstimulation in poor responders. DESIGN: Randomized, controlled study. SETTING: Private center for assisted reproduction. PATIENT(S): One hundred twenty-nine women who were poor responders in a previous IVF cycle. INTERVENTION(S): Fifty-nine women underwent 114 attempts of natural-cycle IVF, and 70 women underwent 101 attempts of IVF with controlled ovarian hyperstimulation with microdose GnRH analog flare. MAIN OUTCOME MEASURE(S): Number of oocytes retrieved, pregnancy rate (PR) per cycle, PR per transfer, and implantation rate. RESULT(S): The poor responders treated with natural-cycle IVF and those treated with micro-GnRH analog flare showed similar PRs per cycle and per transfer. The women treated with natural-cycle IVF showed a statistically significant higher implantation rate (14.9%) compared with controls (5.5%). When subdivided into three groups according to age (or=36-39 years, >or=40 years), younger patients had a better PR than the other two groups. CONCLUSION(S): In poor responders, natural-cycle IVF is at least as effective as controlled ovarian hyperstimulation, especially in younger patients, with a better implantation rate.

Number of embryos transferred and implantation.

Multiple pregnancies are considered the most frequent and serious complication of assisted reproduction technology. To reduce the frequency of multiple pregnancies, several centers have adopted a policy of reducing the number of embryos transferred in the uterus, suggesting single embryo transfer. Even though a significant number of papers have been published on this issue, no general consensus exists on how many embryos to replace in the uterus and at which cleavage stage. We conducted a retrospective study on cycles performed throughout 2003, analyzing the relation between the number of embryos transferred and the pregnancy and implantation rates, evaluating also the role of the woman's age. No differences were found among the groups except in one-embryo transferred women, which were mostly natural cycles, for estradiol levels, number of mature oocytes retrieved, number of top quality embryos, and pregnancy rate. The implantation rate was significantly higher in the two-embryo transfers versus three-embryo transfers. We found higher pregnancy and implantation rates with similar multiple pregnancy rates in patients where only two embryos were transferred versus three embryos transfer when women were less than 35 years old. In women aged less than 35 years, which in turn have the higher expectancy of successful pregnancy and also the higher risk of multiple pregnancy, the single embryo transfer is a suitable choice for these patients.