A novel embryo quality scoring system to compare groups of embryos at different developmental stages

PurposeTo construct a new embryonic quality scoring system to compare groups of embryos at different developmental stages.MethodsBased on a hypothesis that the implantation potential of any embryo in an ovum pickup (OPU) cycle remains the same at any stage of development, be it day 2, 3, or 5, a new embryo quality scoring (EQS) system was designed. It was based on the analysis of the clinical results of 1610 single embryo transfers. We validated this scoring system in the comparison of embryonic quality between groups by evaluating the mean scores calculated at day 2, day 3, and day 5 for 957 embryos (150 cycles) from 3 different groups. We then compared EQSs of patients with pregnancy favorable factors (group A) such as young age and high AMH levels, with the patients with contra features (group B).ResultsWe confirmed that each mean EQS assessed at different stages of embryonic development within the same group was similar. The mean EQSs on day 3 and day 5 in group A were significantly higher than the mean EQSs on days 2, 3, and 5 in group B.ConclusionThe novel EQS system proposed by us enables embryonic quality comparison between groups of embryos at different developmental stages.     

体外受精-胚胎移植周期中不同天数和发育阶段囊胚移植妊娠结局的比较

目的:寻找体外受精-胚胎移植(IVF-ET)治疗中选择合适时期和高质量囊胚进行移植的依据。方法:回顾性分析行囊胚移植的4 237例患者的临床资料,其中新鲜单囊胚移植1 574例,冻融单囊胚移植854例,新鲜双囊胚移植135例,冻融双囊胚移植1 674例。根据囊胚发育天数和发育阶段分为第5日(D5)早期组、D5扩张组、第6日(D6)早期组和D6扩张组。比较各组临床妊娠率、种植率、妊娠结局和新生儿情况等各项指标。结果:①单囊胚移植:新鲜周期D5扩张组临床妊娠率和种植率显著高于其它组,其活产率显著高于D6早期组和D6扩张组,流产率明显低于D6扩张组(P0.05);冻融周期D5扩张组具有较高的复苏率、临床妊娠率和种植率显著高于D5早期组和D6早期组(P0.05);②双囊胚移植:D5移植2枚扩张期囊胚的种植率显著高于D5移植2枚早期囊胚,其他各组间差异无统计学意义(P0.05);冻融周期D5扩张组移植2枚囊胚种植率最高,且临床妊娠率显著高于D5早期组,而流产率明显低于后者,差异有统计学意义(P0.05)。结论:对于单囊胚移植和双囊胚移植,无论新鲜周期或冻融周期,D5扩张组囊胚妊娠结局最佳。     

Coculture of mouse embryos with cryopreserved human oviduct epithelial cells

Purpose The effect of coculturing mouse embryos with cryopreserved human oviduct epithelial cells was investigated. The cryopreserved cells in Cellbanker were thawed and cultured in Richard D. Goldsby culture medium to establish monolayers. Two-cell-stage mouse embryos were cultured alone (control group) or cocultured with monolayers established from cryopreserved cells (cryopreserved coculture group) or from fresh cells (fresh coculture group). The rates of embryo development and the qualities of the blastocysts in the three groups were compared.Results The two coculture groups had significantly higher blastocyst development rates (cryopreserved coculture group, 81.6%; fresh coculture group, 82.2%) than the control group (63.1%). The two coculture groups had significantly more blastomeres (cryopreserved coculture group, 108.3 ± 25.9; fresh coculture group, 108.4 ± 25.1) than the control group (87.7 ± 31.9).Conclusion The method of cryopreservation of human oviduct epithelial cells using Cellbanker is simpler than conventional cryopreservation methods. These cryopreserved human oviduct epithelial cells may provide a constant supply of cells for coculture for in vitro fertilization and embryo transfer.     

Dizygotic twin delivery following in vitro fertilization and transfer of thawed blastocysts cryopreserved at day 6 and 7

OBJECTIVE: To report the first conception and delivery following transfer of thawed human blastocysts maintained in extended in vitro culture with cryopreservation at day 6 and 7. DESIGN: Case report. SETTING: Major urban infertility referral center. PATIENT(S): A 26-year-old woman with pelvic endometriosis and two prior unsuccessful in vitro fertilization/embryo transfer (IVF-ET) attempts. INTERVENTION(S): The patient underwent controlled ovarian hyperstimulation using a combined FSH + hMG protocol, and 24 oocytes were retrieved. MAIN OUTCOME MEASURE(S): Dizygotic twin delivery after IVF and intracytoplasmic sperm injection (ICSI), assisted embryo hatching, and ultrasound-guided transfer of cryopreserved blastocysts. RESULT(S): After three embryos were subjected to assisted hatching, they were transferred fresh on day 3, but no implantation occurred. All nontransferred embryos (n = 11) were observed during extended in vitro culture and three blastocysts were selected for cryopreservation on day 6 and 7; thaw and transfer occurred the following month and a pregnancy was achieved. Dizygotic twins (female/female) were delivered by cesarean in the early third trimester. CONCLUSION(S): Substantial advancements have been made in the field of embryo cryogenics and in vitro fertilization, but controversy remains regarding the value of freezing late-developing human blastocysts. Here we describe the first reported live births with IVF after extended in vitro culture and cryopreservation at day 6 and 7 after fertilization.     

Influence of post-thaw culture on the developmental potential of human frozen embryos

Purpose

Apart from freezing/thawing related cryodamage, several additional factors have been identified as major players in the reduction of success rates after frozen embryo transfers. The post-thaw culture is particularly relevant as it may amplify environmental influences over a stressed embryo. In the present study the influence of the post-thaw culture duration on the implantation and developmental potential of cleavage stage embryos was evaluated.

Methods

In this retrospective evaluation, that spanned an 8-year period, 631 frozen-thawed embryos were allocated to one of two study groups, depending on their post-thaw culture period: 1) the long (18–24 h), or 2) the short (2–5 h) culture group. Groups were compared regarding implantation rate and live birth rate per embryo transferred. This comparison was corrected for the most common confounding factors such as maternal age at oocyte pick-up, number of transferred embryos, developmental day at freezing, blastomere survival after thawing, catheter used for transfer and year of procedure.

Results

Implantation and live birth rate per embryo transferred were inversely related to the duration of the post-thaw culture, as diminishing this period significantly increased both rates. Moreover, no advantage could be found for a long post-thaw culture period, even for embryos with observed mitotic activity.

Conclusion

This retrospective analysis indicates that a short post-thaw culture period is associated with higher implantation and live birth rates per embryo. This study supports selection of frozen-thawed embryos strictly based on blastomere cryosurvival and raises the hypothesis that environmental factors may have an important role on embryo implantation and developmental potential during post-thaw culture.     

Initial experience with extended culture and blastocyst transfer of cryopreserved embryos

Objective: Our purpose was to evaluate the viability and transfer efficiency of cryopreserved embryos allowed to develop into blastocysts in extended culture for in vitro fertilization. Study Design: The embryos for in vitro fertilization that had been cryopreserved at either 2 PN (pronuclear) or cleaving stage (day 1-3) were thawed and cultured for uterine transfer on day 5. Outcome for day 5 embryo transfer was prospectively compared with previous outcomes from embryos transferred on day 2 or 3. Results: For embryos thawed and transferred on day 2 or 3 (n = 99), the pregnancy rate was 33%, the implantation rate per embryo transferred was 15.2%, and the rate of multiple gestations was 42.4% (14/33) with 35.7% of pregnancies having ≥3 gestational sacs. For extended culture embryos transferred on day 5 (n = 25), the pregnancy rate was 36%, the implantation rate per embryo transferred was 16.7%, and the rate of multiple gestations was 33.3% (3/9) with all of these being twins. For embryo transfers performed on day 5 in which only blastocysts were transferred (n = 9), the pregnancy rate was 66.7%, the implantation rate per blastocyst was 44.4% (greater than the rate for the day 2 or 3 embryos, P = .0043), and the rate of multiple gestations was 33.3% (2/6) with all of these being twins. In extended culture 29.8% of cryopreserved embryos progressed to the blastocyst stage. In this series 4 subjects (15.4%) did not have blastocysts by day 5. Conclusion: Acceptable pregnancy rates can be obtained from cryopreserved embryos cultured to the blastocyst stage with a significantly higher implantation rate. Transfer of embryos that have “self-selected” to blastocysts results in reduced risk of higher-order (>2) multiple gestations, because only 1 or 2 embryos are transferred. (Am J Obstet Gynecol 1999;180:1472-4.)     

Cryopreservation of human embryos at the morula stage and outcomes after transfer

OBJECTIVE: To evaluate the survival rate of human morula embryo freezing and the morphological alterations during freezing, during and after thawing, and their applications in embryo selection. DESIGN: Retrospective observational study. SETTING: Private infertility clinic. PATIENT(S): Consecutive patients under age 39 undergoing frozen morula embryo transfers from December 1999 to May 2003. INTERVENTION(S): Embryo freezing was performed at the morula stage. Embryo thaw and post-thaw ETs were conducted on the same day, which is equivalent to a day 4 ET. MAIN OUTCOME MEASURE(S): Morphological alterations during freezing and thawing and after thawing. Post-thaw embryo survival rates, transferable rates, pregnancy rates, and implantation rates. RESULT(S): Morula embryos showed reversed morphological alterations during the freezing process; these alterations were recovered during thawing or shortly after the thawing. Post-thaw survival rates showed no significant difference between any of the morula substages. However, embryos scored as grade 3, which represented good quality, had significantly higher post-thaw survival and transferable rates than grade 2 and 1 embryos. Patients who received at least one grade 3 embryo had significantly higher pregnancy rates, implantation rates, and ongoing/live birth rates than other groups. CONCLUSION(S): An acceptable survival rate can be achieved after cryopreservation of human morula embryos, and morphological alterations that occur during and shortly after an embryo thaw can be a feasible index for determining viable embryos.     

妊娠与非妊娠女性卵巢肿瘤蒂扭转的临床特点及诊治比较#br#

Objective?To explore the clinical characteristics, diagnosis and treatment of patients with ovarian tumor pedicle torsion in pregnancy and non-pregnancy. Methods?A retrospective analysis of the general, clinical, surgical and pathological data of 163 patients who were diagnosed with ovarian tumor pedicle torsion after surgery were admitted to the First Affiliated Hospital of Xi'an Jiaotong University from January 2009 to December 2018. It is divided into pregnant group and non-pregnant group. Results?(1) The incidence of abdominal pain, nausea and vomiting, and peritoneal irritation in the pregnant group were lower than those in the non-pregnant group, but the differences were not statistically significant (all P>0.05). (2) The interval between the symptoms in the pregnancy group and the hospital stay was longer than that in the non-pregnancy group, and the difference was statistically significant (P<0.05). The interval between hospitalization and surgery was significantly shorter in the pregnant group than in the non-pregnant group (P<0.05). (3) The average diameter of cysts in the pregnant group was smaller than that in the non-pregnant group, and the difference was statistically significant (P<0.05). (4) In non-pregnancy group, the most common postoperative pathological type was teratomas in 34 cases (26.8%). In pregnancy group, the most common type was physiological cysts in 10 cases (27.8%). (5) In pregnancy group, 4 patients with early pregnancy required simultaneous abortion. A 23-week pregnant woman presented with abdominal pain 1 week later and had a spontaneous abortion 4 hours after surgery; Twenty-six cases were followed up to term, and the pregnancy outcome was good; 5 cases were lost to follow-up. Conclusion?The clinical symptoms of ovarian tumor pedicle torsion during pregnancy are not specific, and it is easy to delay diagnosis and treatment. Adnexal surgery did not increase miscarriage or adverse pregnancy outcomes.     

Clinical outcome of day 2 versus day 5 transfer in cycles with one or two developed embryos

OBJECTIVE: To determine whether extended culture of embryos to blastocysts has any benefit in cycles with only one or two created embryos. DESIGN: Retrospective analysis of cycles comparing outcomes of day 2 and day 5 transfers. Our day 2 group was from the year 1999 and our day 5 group, from the year 2000. SETTING: Assisted reproductive technology program of a teaching hospital. PATIENT(S): All patients, irrespective of age, who had developed one or two embryos. INTERVENTION(S): Stimulated IVF, intracytoplasmic sperm injection, or testicular sperm extraction and intracytoplasmic sperm injection cycles with 2-day culture in universal IVF medium (n = 133) or 5-day culture in BlastAssist media (MediCult, Jyllinge, Denmark; n = 132). MAIN OUTCOME MEASURE(S): Pregnancy, implantation, and take-home baby rates. RESULT(S): In the groups of 2-day and 5-day culture, embryo transfer was performed in 98% and in 57% of cycles, respectively. However, the total implantation rate per created embryo (18% vs. 18%), the pregnancy rate per cycle (23% vs. 21%), and the take-home baby rate (69.4% vs. 71.4%) did not differ between the day 2 and day 5 groups. CONCLUSION(S): Extended culture of embryos does not improve or decrease their capacity for implantation but only allows for better selection and is therefore not necessary in cycles with fewer than three embryos.     

Development of spare human preimplantation embryos in vitro: An analysis of the correlations among gross morphology,cleavage rates,and development to the blastocyst

Following in vitro fertilization, the criteria commonly used to select human embryos for transfer are the cleavage rate and gross morphology, the contention being that those embryos which divide more rapidly and have regular, spherical blastomeres are more likely to lead to a pregnancy. In order to assess the validity of this assumption, the development in vitro of spare embryos was investigated. Eggs and embryos were cultured in Earle's balanced salt solution containing 10% heat-inactivated patient's serum, and insemination was performed at 40 hr post human chorionic gonadotropin (hCG). At 82–90 hr post hCG, up to four embryos were transferred. Any spare embryos were cultured in the same medium for up to 6 days and scored daily for cell number and morphology using a quality scale of 4-1 according to degree of fragmentation and shape of the blastomeres. Of 317 fertilized eggs, 55 (17%) developed to the fully expanded blastocyst stage. The remaining embryos ceased development at the one-cell (6; 2%), two-cell (49; 15%), fourcell (110; 35%), eight-cell (61; 19%), and cavitating morula (36; 11%) stages. The relationship between developmental arrest and gross morphology is discussed.     

Outcome of frozen embryo replacement cycles following elective cryopreservation of all embryos in women at risk of developing ovarian hyperstimulation syndrome

Aim: Our aim was to compare the outcome in subsequent frozen embryo replacement cycles in four groups of patients who had elective cryopreservation of all their embryos because they were considered to be at increased risk of developing severe ovarian hyperstimulation syndrome. Design: Sixty-two (91%) of 68 IVF cycles (68 patients) in which elective cryopreservation of all embryos was performed were analyzed. All patients continued on the GnRH agonist, buserelin, after oocyte recovery until the onset of vaginal bleeding. Frozen embryo replacement occurred in a hormone replacement cycle that started either on day 3 of the withdrawal bleed (group I;N=15) or after serum estradiol levels had fallen to <100 pmol/L (group II;N=16). The other patients commenced a frozen embryo replacement cycle several months later in either a hormone replacement (group III;N=15) or a natural (group IV;N=16) cycle. Results: Two patients developed severe ovarian hyperstimulation syndrome. There were no significant differences among the four groups regarding demographic variables, the dose of hMG used, and the clinical outcome. There was a higher but not significantly different clinical pregnancy rate in group I (26.7%), compared to group II (12.5%), group III (13.3%), and group IV (18.8%). Conclusions: Several options exist for the timing and protocol used for frozen embryo replacement in patients who had elective cryopreservation for the prevention of ovarian hyperstimulation syndrome, none of which was found to be clearly superior in this observational report.Presented at the 1994 Annual Conference of the American Fertility Society.     

Clinical experience and perinatal outcome of blastocyst transfer after coculture of human embryos with human endometrial epithelial cells: a 5-year follow-up study

OBJECTIVE: To evaluate the reproductive and neonatal outcome of blastocyst transfer after coculture with human endometrial epithelial cells in IVF and oocyte donation. DESIGN: Retrospective study.Private assisted reproductive center. PATIENTS(S): Two hundred sixty women undergoing IVF and 469 oocyte recipients. INTERVENTION(S): IVF or intracytoplasmic sperm injection (ICSI) and transfer of at least one blastocyst after coculture with human endometrial epithelial cells. MAIN OUTCOME MEASURE(S): Blastocyst formation rate, implantation and pregnancy rates, neonatal outcome, and congenital birth defects. RESULT(S): Among patients who had transfer with their own oocytes, 1193 of 2349 cocultured embryos developed up to the blastocyst stage (50.8%), and pregnancy and implantation rates of 33.9% and 19.2%, respectively, were achieved. In the oocyte donation program, 1819 blastocysts were obtained from 3127 embryos (58.2%), with subsequent pregnancy and implantation rates of 57.0% and 31.0%, respectively. The blastocyst rate remained stable throughout the 5 years of the study, but the pregnancy and implantation rates increased dramatically. Of 139 deliveries, 57 (41.0%) were multiple pregnancies and 1 (0.7%) was a multifetal birth (four live born infants). Out of 200 children born, 59% were male, and congenital birth defects were observed in 2.5%. CONCLUSION(S): Coculture of human embryos with endometrial epithelial cells yields a blastocyst formation rate of 50.8% to 58.2% and encouraging implantation and pregnancy rates. This technique reduces the mean number of embryos transferred in each patient. The number of embryos implanted is more relevant to neonatal outcome than is the coculture system and blastocyst transfer used. The risk of congenital birth defects associated with this program is similar to that recorded in early ET in IVF or ICSI.     

全部胚胎冷冻保存在卵巢低反应患者微刺激方案IVF-ET中的应用

目的:探讨全部胚胎冷冻保存对卵巢低反应患者微刺激方案助孕结局的影响。方法:回顾性分析微刺激方案IVF-ET共483个周期,根据胚胎移植时机不同分为A组(新鲜胚胎移植组,275个周期)和B组[全部胚胎冷冻保存首次冷冻胚胎复苏移植术(FET)组,208个周期],比较组间患者的年龄、基础FSH、不孕年限、扳机日优势卵泡数目、内膜厚度、获卵数、可利用胚胎数及临床妊娠率、胚胎种植率、流产率。结果:A组扳机日内膜厚度和获卵数高于B组,差异有统计学意义(P0.05),A组的临床妊娠率、胚胎种植率低于B组,差异有统计学意义(P0.05),患者的年龄、基础FSH、Gn总量、扳机日优势卵泡数、扳机日雌二醇(E2)和孕酮(P)值、可利用胚胎数、移植周期内膜厚度、移植胚胎数、流产率组间均无统计学差异(P0.05)。结论:对于使用微刺激方案助孕的卵巢低反应患者,全部胚胎冻存择期进行FET可以改善其助孕结局,是一种值得临床推广的助孕策略。