Luteal Phase Support Using Subcutaneous Progesterone: A Systematic Review

Luteal phase support (LPS) is crucial in assisted reproductive technology (ART) cycles when the luteal phase has been found to be defective. Such deficiency is most likely related to the supraphysiological steroid levels that usually occurr in stimulated cycles which, in turn, could severely affect luteinizing hormone (LH) secretion and function, thereby negatively influencing the luteal phase. A number of different medications and routes have been successfully used for LPS in ART. Although an optimal protocol has not yet been identified, the existing plethora of medications offer the opportunity to personalize LPS according to individual needs. Subcutaneous administration progesterone has been proposed for LPS and could represent an alternative to a vaginal and intramuscular route. The aim of the present systematic review is to summarize the evidence found in the literature concerning the application of subcutaneous progesterone in ARTs, highlighting the benefits and limits of this novel strategy. With this aim in mind, we carried out systematic research in the Medline, ISI Web of Knowledge, and Embase databases from their inception through to November 2020. Randomized controlled trials (RCTs) were preferred by the authors in the elaboration of this article, although case-control and cohort studies have also been considered. According to our findings, evidence exists which supports that, in women with a good prognosis undergoing a fresh in vitro fertilization (IVF) cycle, subcutaneous Pg is not inferior to vaginal products. In the Frozen-thawed embryo transfer (FET) cycle, data concerning efficacy is mixed with an increased miscarriage rate in women undergoing a subcutaneous route in oocyte donor recipients. Data concerning the acceptance of the subcutaneous route versus the vaginal route are encouraging despite the different scales and questionnaires which were used. In addition, a cost-effective analysis has not yet been conducted

Endometrial microbiota profile in in-vitro fertilization (IVF) patients by culturomics-based analysis

IntroductionIt is well recognized that the human uterus and adjoining tissues of the female reproductive tract exist in a non-sterile state where dysbiosis can impact reproductive outcomes. The endometrial microbiota is a part of this greater milieu. To date, it has largely been studied using 16S rRNA or metagenomics-based methodologies. Despite the known advantages of sequencing analysis, several difficulties have been noted including sample contamination and standardization of DNA extraction or sequencing. The aim of this study was to use a culturomics-based method to analyze the endometrial microbiota and correlate the results with ongoing pregnancy rates.MethodsA prospective cohort study was performed at the University of Naples from June 2022 to December 2022. Ninety-three patients undergoing an IVF cycle with single embryo transfer (ET) (fresh or frozen) were enrolled in the study. Following ET, the catheter tip was inserted into brain heart infusion (BHI) medium under sterile conditions for culture. After 24h and 48h of incubation the microorganisms in the colonies were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).ResultsOverall, 68 (73,92%) patients resulted positive for one or more microbes and 25 patients (26,08%) had no microbial growth. Across all participants, the four most important phyla were Firmicutes (87,76%), Proteobacteria (27,94%), Actinobacteria (10,29%) and Ascomycota (8,82%). Lactobacillus species, in particular, was significantly correlated with ongoing pregnancy rate (p=0,05). On the other hand, Staphylococcus subspecies (spp.) (p<0,05) and Enterobacteriaceae (p<0,001) were found to have a negative impact on the implantation rate.DiscussionDetection of bacteria by culturomics from catheter tips used for embryo transfer has been shown to be a reliable method to detect pathogen growth. Endometrial microbiota testing in clinical practice could certainly offer a means to further improve diagnosis and treatment strategies in IVF patients

Differences in amniotic amino acid concentrations between pregnancies obtained with transfer of vitrified thawed <i>in vitro</i>–produced embryos and with natural mating in sheep

In vitro embryo production (IVP) and cryopreservation are associated with a high incidence of pregnancy complications and fetal abnormalities that may be linked with alterations of placental development. The amniotic fluid is partly derived from the transport of water and solutes across the placenta and provides the fetus with amino acids (AAs), which are the building blocks for biomolecules involved in physiological growth and development. To better understand the anomalies associated with IVP pregnancies, the present study was conducted to test the hypothesis that amniotic concentrations of AAs differ in pregnancies derived from vitrified/thawed (V/T) IVP embryos compared with gestations obtained with natural mating (NM) in sheep. Amniotic fluid was sampled in ewes that were pregnant after transfer of V/T IVP embryos and that had conceived with NM between Days 60 and 65 (V/T, n = 6; NM, n = 11) and between Days 80 and 85 (V/T, n = 5; NM, n = 14) of gestation via ultrasound-guided amniocentesis. Concentrations of 16 AAs in the amniotic fluid were measured using high-performance liquid chromatography. From Days 60 to 65 of gestation, concentrations of cystine, phenylalanine, and isoleucine were lower in V/T compared with NM ewes. From Days 80 to 85 of pregnancy, the mean concentrations of cystine and lysine were lower in the V/T versus NM groups. The total AA concentration per ewe was similar between the groups from Days 60 to 65 and 80 to 85 of gestation and decreased by 55% from Days 60 to 65 and 80 to 85 of gestation in all ewes. The most abundant AA from Days 60 to 65 of gestation was alanine in both groups, whereas from Days 80 to 85, the most abundant AAs were alanine in NM and glycine in V/T ewes; cystine was the less abundant detectable AA in all ewes at both stages of gestation. Results report that V/T IVP embryos have decreased concentrations of individual AAs in the amniotic fluid during the second trimester of gestation possibly because of an impaired placental vasculogenesis or because of a reduced placental transport. These novel findings are relevant to unravel the mechanisms responsible for the issues of pregnancies achieved with the transfer of IVP and cryopreserved embryos

Impact of polymorphisms of gonadotropins and their receptors on controlled ovarian stimulation: a prospective observational study

Study question: Which effect do polymorphisms of gonadotropins and their re- ceptors have on stimulation outcomes in IVF patients co-treated with a GnRHa long down-regulation protocol? Summary answer: Allele C of FSHR-29, LHCGR-291 and FSHR-680 all resulted in a significantly increased cumulative r-FSH dose: total number of oocytes or mature oocytes ratio. What is known already: Specific polymorphisms might influence controlled ovarian stimulation in women undergoing IVF/ICSI. Data regarding the pos- sible interactions of these polymorphisms are still scanty, especially as regards LHCG-R polymorphisms. Study design, size, duration: Prospective observational study in 100 normogo- nadotropic IVF/ICSI patients came from three public IVF Units. Participants/materials, setting, methods: Normogonadotropic Caucasian women fulfilling the following inclusion criteria were enrolled: age 20–34 years; BMI 20–27 kg/m2; basal FSH ≤ 10 IU/l; functional ovaries. Exclusion crite- ria were: uterine anomalies; endocrine, genetic or immunological disorders; PCOS; history of impaired ovarian response (≤ 4 oocytes retrieved) in at least one IVF/ICSI cycle. Patients underwent a GnRH long down-regulation protocol with a starting dose of 150 IU of recombinant FSH daily. Six polymorphisms were genotyped. Main results and the role of chance: The following polymorphisms were analyzed: FSHR-680 (rs6166); FSHR-min29 (rs1394205); LHCGR intronic (rs4073366); LHCGR-291 (rs 12470652); LHCGR-312 (rs2293275); FSHβ- 2623 (rs6169). Basal FSH levels were significantly lower in homozygotic carriers of FSHR-630 (T/T) than in heterozygotic C/T (p = 0.023). Lower basal estradiol levels were seen in homozygotic carriers of FSHR-29 promoter C/C compared to heterozygotic C/T (p = 0.045). Basal estradiol levels and number of fertil- ized and mature oocytes were lower in homozygotic carriers of LHCGR-291 (T/T) compared to heterozygotic C/T (p = 0.035 and p = 0.05 respectively). The presence of allele C on both FSHR-min29 and LHCGR-291 caused an in- creased ratio between the cumulative r-FSH consumption and the total number of oocytes as well as mature oocytes (RR: 5.47, CI 95%: 3.13–7.81, p < 0.001). This observation was also confirmed when polymorphisms of FSHR-680 were included in the analysis. Specifically, the presence of allele C on these three genes was related to an increased ratio between the cumulative FSH consump- tion and the total number of oocytes or mature oocytes (RR: 5.44, CI 95%: 3.18–7.71, p < 0.001). Limitations, reasons for caution: Although limited by the small size of the population, these findings confirm a possible interaction between multiple poly- morphisms in assisted reproductive technology. Wider implications of the findings: These data support the concept that the ovarian response to exogenous FSH seems to be determined by the in- teraction of specific genetic traits. Moreover, this study shows an involve- ment of the LHCGR-291 polymorphism in ovarian response to exogenous gonadotropins. Trial registration number: Not applicable

Post‐mortem recovery, in vitro maturation and fertilization of fallow deer (Dama dama, Linnaeus 1758) oocytes collected during reproductive and no reproductive season

Habitat degradation leads to small and fragmented populations, lower genetic variability and fertility overtime. Assisted reproductive techniques represent important tools to cope with the dramatic loss of biodiversity. Fallow deer (Dama dama), beyond its high commercial value and wide distribution, may represent the most suitable model to study endangered cervids. In this study, oocytes were recovered post‐mortem from fallow deer during the breeding and no breeding seasons and were in vitro matured (IVM). The ability of cryopreserved thawed sperm samples recovered by electroejaculation from four adult males was tested by in vitro fertilization of IVM oocytes. The number of oocytes collected per ovary did significantly vary across seasons from 6.2 ± 0.92 during breeding season to 10.4 ± 1.26 during no breeding season (p = .006). Oocytes collected during the breeding season showed higher in vitro fertilization rate compared to the no breeding season (p = .045). However, no embryos reached the blastocyst stage. Semen samples obtained by electroejaculation were successfully cryopreserved, although the cryopreservation process negatively affected most kinetic parameters, mainly at 2 hr post‐thawing. Moreover, the percentage of rapid spermatozoa significantly decreased between fresh samples and at 2 hr post‐thawing, whereas the percentage of slow spermatozoa increased across the same period (p < .05). Our study provides the logistic steps for the application of assisted reproductive techniques in fallow deer and might be of great interest for genetic resource bank planning.Peer reviewe