P–643 Is Euploid blastocyst number higher in luteal versus follicular phase? A case-control study of IVF outcomes of follicular versus luteal phase ovarian stimulation

Study question Is there a difference between IVF outcomes in patients undergoing follicular versus luteal phase ovarian stimulation in different menstrual cycles? Summary answer Number of euploid blastocyst were higher in luteal phase ovarian stimulation IVF cycles. All other outcomes were similar between follicular and luteal phase IVF cycles. What is known already It has been published that human beings can have two or three follicular recruitment waves as observed in animals studies a long time ago. From these findings, several recent studies showed that two egg retrievals at the same menstrual cycle, named as Duo Stim, optimize time and IVF outcomes in women with low ovarian reserve due to more eggs retrieved in a shorter period with consequently higher probability of having good embryos to transfer. However, there is no knowledge about diferences concerning IVF outcomes between folicular and luteal ovarian stimulation, performed at the same women in different menstrual cycles. Study design, size, duration Retrospective, case-control study in a single IVF center. One-hundred-two patients who had two IVF treatments – the first cycle initiating ovarian stimulation at follicular phase (FPS) and the second cycle initiating after a spontaneous ovulation at luteal phase (LPS) – in different menstrual cycles (until 6 months apart) between 2014 and 2020, were included. Statistical analysis was performed with Mann-Whitney test and was considered significant when p ≤ 0.05. Data is represented as mean±SD. Participants/materials, setting, methods Patients underwent two IVF treatments in different menstrual cycles; the FPS IVF treatment was initiating at D2/D3 of menstrual cycle and the LPS treatment started three or four days after spontaneous ovulation, if at least 4 antral follicles were detected. Both IVF treatments were performed with and antagonist protocol and freeze all strategy. The majority of patients presents low ovarian reserve/Ovarian age as primary infertility factor (84.3%). Main results and the role of chance Patient’s mean age was 39.30±3.15 years, BMI (22.66±3.16) and AMH levels (0.85±0.85 ng/mL). Comparison of hormonal levels at the beginning of ovarian stimulation showed differences for FPS vs LPS, as expected: E2 (39.69±31,10 pg/mL vs 177.33±214.26 pg/mL,p< 0.0001) and P4 (0.76±2.47ng/mL vs 3,00±5.00 ng/mL,p< 0.0001). However, E2 and P4 at the day of oocyte maturation trigger were not different between FPS and LPS (1355.24±895.73 pg/mL vs 1133.14±973.01 ng/mL,p=0.0883 and 1.12±1.49 ng/mL vs 2.94±6.51,p=0.0972 respectively). There was no difference for total dose of gonadotrofins (FPS 2786.43±1102.39.01UI vs LPS 2824.12±1188.87UI, p = 0,8578), FSH (FPS 9.50±4.98 vs LPS 11.90±12.99,p=0.7502) and AFC (FPS 7.13±4.25 vs LPS 6.42±4.65,p=0,0944). From 102 patients that started ovarian stimulation, 78 had 1 or more oocyte collect in FPS group and 75 in LPS group: OPU (FPS 4.78±4.93 vs LPS 4.65±5.54,p=0.7889), number of MII (FPS 3.21±3.52 vs LPS 3.40±4.53,p=0.7889). From those, 52 patients performed ICSI in both cycles; fertilization rate 64.9%±28.6% for FPS vs 62.1%±32.4% for LPS,p=0.7899) and blastocyst formation 2.15±2.15 for FPS vs 2.54±2.35,p=0.3496). Data from 25 patients who had embryo biopsy for PGT-A showed similar number of blastocyst biopsed (2.12±1.72 FPS vs 2.48±1.71 LPS,p=0.3101) and a statistically significant difference regarding number of euploid blastocyst (0,20±0,41 FPS vs 0,96±0,93 LPS,p=0,0008). Limitations, reasons for caution This is a retrospective study in a limited number of patients. Therefore, it is not possible to make a definitive conclusion that LPS proportionate higher number of euploid than FPS. More studies are necessary to investigate not only IVF outcomes but also the impact on pregnancy rates. Wider implications of the findings: In our study, LPS protocol after spontaneous ovulation, presents similar IVF outcomes compared to routinely FPS protocol. Intriguingly, the number of euploid blastocyst was significant higher in LPS, which may be further investigated. In this way, LPS is another option of IVF treatment, and may optimize time and treatment results. Trial registration number Not applicable

P-643 Pre-selected for an award: Is Euploid blastocyst number higher in luteal versus follicular phase? A case-control study of IVF outcomes of follicular versus luteal phase ovarian stimulation

Study question Is there a difference between IVF outcomes in patients undergoing follicular versus luteal phase ovarian stimulation in different menstrual cycles? Summary answer Number of euploid blastocyst were higher in luteal phase ovarian stimulation IVF cycles. All other outcomes were similar between follicular and luteal phase IVF cycles. What is known already It has been published that human beings can have two or three follicular recruitment waves as observed in animals studies a long time ago. From these findings, several recent studies showed that two egg retrievals at the same menstrual cycle, named as Duo Stim, optimize time and IVF outcomes in women with low ovarian reserve due to more eggs retrieved in a shorter period with consequently higher probability of having good embryos to transfer. However, there is no knowledge about diferences concerning IVF outcomes between folicular and luteal ovarian stimulation, performed at the same women in different menstrual cycles. Study design, size, duration Retrospective, case-control study in a single IVF center. One-hundred-two patients who had two IVF treatments – the first cycle initiating ovarian stimulation at follicular phase (FPS) and the second cycle initiating after a spontaneous ovulation at luteal phase (LPS) – in different menstrual cycles (until 6 months apart) between 2014 and 2020, were included. Statistical analysis was performed with Mann-Whitney test and was considered significant when p ≤ 0.05. Data is represented as mean±SD. Participants/materials, setting, methods Patients underwent two IVF treatments in different menstrual cycles; the FPS IVF treatment was initiating at D2/D3 of menstrual cycle and the LPS treatment started three or four days after spontaneous ovulation, if at least 4 antral follicles were detected. Both IVF treatments were performed with and antagonist protocol and freeze all strategy. The majority of patients presents low ovarian reserve/Ovarian age as primary infertility factor (84.3%). Main results and the role of chance Patient’s mean age was 39.30±3.15 years, BMI (22.66±3.16) and AMH levels (0.85±0.85 ng/mL). Comparison of hormonal levels at the beginning of ovarian stimulation showed differences for FPS vs LPS, as expected: E2 (39.69±31,10 pg/mL vs 177.33±214.26 pg/mL, p < 0.0001) and P4 (0.76±2.47ng/mL vs 3,00±5.00 ng/mL,p < 0.0001). However, E2 and P4 at the day of oocyte maturation trigger were not different between FPS and LPS (1355.24±895.73 pg/mL vs 1133.14±973.01 ng/mL,p = 0.0883 and 1.12±1.49 ng/mL vs 2.94±6.51,p = 0.0972 respectively). There was no difference for total dose of gonadotrofins (FPS 2786.43±1102.39.01UI vs LPS 2824.12±1188.87UI, p = 0,8578), FSH (FPS 9.50±4.98 vs LPS 11.90±12.99, p = 0.7502) and AFC (FPS 7.13±4.25 vs LPS 6.42±4.65,p = 0,0944). From 102 patients that started ovarian stimulation, 78 had 1 or more oocyte collect in FPS group and 75 in LPS group: OPU (FPS 4.78±4.93 vs LPS 4.65±5.54,p = 0.7889), number of MII (FPS 3.21±3.52 vs LPS 3.40±4.53,p = 0.7889). From those, 52 patients performed ICSI in both cycles; fertilization rate 64.9%±28.6% for FPS vs 62.1%±32.4% for LPS,p = 0.7899) and blastocyst formation 2.15±2.15 for FPS vs 2.54±2.35,p = 0.3496). Data from 25 patients who had embryo biopsy for PGT-A showed similar number of blastocyst biopsed (2.12±1.72 FPS vs 2.48±1.71 LPS,p = 0.3101) and a statistically significant difference regarding number of euploid blastocyst (0,20±0,41 FPS vs 0,96±0,93 LPS,p = 0,0008). Limitations, reasons for caution This is a retrospective study in a limited number of patients. Therefore, it is not possible to make a definitive conclusion that LPS proportionate higher number of euploid than FPS. More studies are necessary to investigate not only IVF outcomes but also the impact on pregnancy rates. Wider implications of the findings In our study, LPS protocol after spontaneous ovulation, presents similar IVF outcomes compared to routinely FPS protocol. Intriguingly, the number of euploid blastocyst was significant higher in LPS, which may be further investigated. In this way, LPS is another option of IVF treatment, and may optimize time and treatment results. Trial registration number Not Applicable

Luteal phase ovarian stimulation versus follicular phase ovarian stimulation results in different human cumulus cells gene expression: a pilot study

OBJECTIVE: Physiologic elevated levels of progesterone in luteal phase can impede early-onset LH surge. However, the impact of high levels of progesterone on the oocyte or cumulus cells (CCs) remains indistinct. Therefore, the aim of study was to investigate the CCs gene expression between luteal phase ovarian stimulation (LPOS) and follicular phase ovarian stimulation (FPOS) in poor ovarian responders (PORs) undergoing in vitro fertilization (IVF) cycles. MATERIALS AND METHODS: This was a prospective non-randomized trial (ClinicalTrials.gov Identifier: NCT03238833). A total of 36 PORs who conformed Bologna criteria and underwent IVF cycles were enrolled. 15 PORs were allocated to the LPOS group and 21 PORs were allocated to the FPOS group. Basic characteristics, cycle characteristics and pregnancy outcomes were compared between the two groups. Moreover, CCs genes regarding inflammation (CXCL1, CXCL3, TNF, PTGES), oxidative-phosphorylation (NDUFB7, NDUFA4L2, SLC25A27), apoptosis (DAPK3, BCL6B) and metabolism (PCK1, LDHC) were analyzed using real-time quantitative PCR between the two groups. RESULTS: Basic characteristics and IVF outcomes were similar between the two groups except significantly high progesterone level in the LPOS group. The mRNA expression of CXCL1 and PTGES were significantly lower in the LPOS group than in the FPOS group ( p < 0.05). The LPOS group had significantly lower mRNA expression of NDUFB7 and NDUFA4L2 than the FPOS group ( p < 0.05). DAPK3 and BCL6B mRNA expression were significantly higher in the LPOS group compared to FPOS group ( p < 0.05). Increased expression of PCK1 and decreased expression of LDHC were observed in the LPOS group compared to the FPOS group. ( p < 0.05). CONCLUSIONS: Compared to the FPOS, the LPOS seemed to reduce favorable inflammation and mitochondrial function, and induce apoptosis and abnormal glucose metabolism in CCs.

Luteal-phase protocol in poor ovarian response: a comparative study with an antagonist protocol

Objective This retrospective study compared the effect of the luteal phase ovarian stimulation protocol (LP group) with the gonadotrophin-releasing hormone (GnRH) antagonist protocol (AN group) in women with poor ovarian responses. Methods Ovarian stimulation was initiated with 225 IU of human gonadotrophin (hMG) daily. When the dominant follicle diameter exceeded 13 mm, 0.25 mg of a GnRH antagonist was used daily until human chorionic gonadotrophin (HCG) administration in the AN group. A GnRH antagonist was not used in the LP group. Ovulation was induced with HCG for all patients when at least one follicle reached a diameter of 16 mm or one dominant follicle reached 18 mm. The highest quality embryos were transferred or cryopreserved for later transfer. Results From January 2013 to December 2015, 274 women with poor ovarian response were included. A total of 108 patients underwent the luteal phase ovarian stimulation protocol while 166 patients underwent the GnRH antagonist protocol. hMG was used for more total days in the LP group was than in the AN group. Oestradiol levels on the day of HCG administration in the LP group were significantly lower than those in the AN group. The mean number of oocytes retrieved in the LP and AN groups was 3.5 ± 2.5 and 3.5 ± 2.9, respectively. The mean number of embryos of the highest quality was 1.7 ± 1.2 and 1.7 ± 1.5, respectively. The clinical pregnancy and implantation rates in the LP and AN groups were 26.2% (22/84) and 25% (29/116), and 15.5% (24/155) and 16.3% (35/215), respectively. Conclusions The luteal phase ovarian stimulation protocol can be applied in women with poor ovarian response and attain comparable clinical pregnancy and implantation rates to those of the GnRH antagonist protocol.