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Purpose: Assisted reproduction technologies (ART) are associated with increased risks of pregnancy complications and obstetric interventions. Here, we aimed to determine if ART affects placental inflammation and oxidative stress as a mechanism for unfavorable pregnancy outcomes., Methods: The levels of six cytokines (IFN-γ, IL-1β, IL-6, IL-8, IL-10, TNFα) were measured using multiplex ELISA. The activity of four antioxidant enzymes (glutathione S-transferase (GST), glutathione peroxidase (GPx), glutathione reductase, superoxide dismutase) and levels of two antioxidants (GSH, vitamin E) were measured using commercial/in-house assays. Markers were compared between ART and unassisted pregnancies, and then groups were stratified using ICD9/10 codes to determine differences in specific clinical contexts., Results: In unassisted twin pregnancies, there was a trend of decreased cytokine levels (IL-1β, IL-6, IL-8, TNFα, p < 0.05), but cytokines in ART twins were the same or higher. Additionally, GST and GPx activities were lower in unassisted twins, and vitamin E levels were higher in ART twins (p < 0.05). In pregnancies complicated by chorioamnionitis, there was a trend of increased cytokine levels in unassisted pregnancies (IL-1β, IL-6, and IL-8, p < 0.05). No increase was observed in ART, and IFN-γ and TNFα were decreased (p < 0.05). Placental GST and GPx activities were higher in unassisted pregnancies with chorioamnionitis compared to ART (p < 0.05)., Conclusion: Attenuation of protective placental inflammatory and oxidative stress responses may play a role in the underlying pathogenesis of negative birth outcomes in ART, expanding our understanding of adverse pregnancy outcomes when ART is used to conceive., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

According to World Health Organization (WHO) reports about 70 million couples suffer from infertility all over the world. A lot of research groups are working on this issue and have made therapeutic approaches by integrating biology, medicine, genetics, chemistry, psychology, mechanic, and many other branches of science. However, these methods have their own pros and cons. Assisted Reproductive Technologies (ART) has appeared to solve infertility problems. In Vitro Fertilization (IVF), Intracytoplasmic Sperm Injection (ICSI), Intrauterine Insemination (IUI) are the most common and conventional technologies in this regard. There are at least two characteristics of microfluidics, mechanical and biochemical, which can be influential in the field of mammalian gamete and preimplantation embryo biology. These microfluidic characteristics can assist in basic biological studies on sperm, oocyte and preimplantation embryo structure, function and environment. Using microfluidics in sorting sperm, conducting different steps of oocyte selection and preparation, and transferring embryo by passing sub-microliter fluid through microchannels results in low cost and short time. The size and shape of microchannels and the volume of used fluid differs from non-human cells to human cells. The most progressions have been seen in animal models. Results suggest that microfluidic systems will lead to improved efficiencies in assisted reproduction., (Copyright © 2020. Published by Elsevier Inc.)

Embryo cryopreservation is a valuable technique in assisted reproductive technology (ART) that increases cumulative pregnancy rates and allows postponement of embryo transfer in patients with undesirable uterine or clinical conditions. Although vitrification has been considered the most efficient method to freeze oocytes and embryos, it is time-consuming and highly operator-dependent. Gavi® is the first semi-automated machine for vitrification capable of controlling crucial variables such as temperature, volume, concentration and exposure time during the vitrification process. We report the first two pregnancies obtained with blastocysts cryopreserved with the Gavi® semi-automated vitrification system in Europe. These outcomes suggest that the utilization of semi-automated vitrification may contribute to improve the outcomes and laboratory logistics of fertility clinics., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

This month's Views and Reviews sets the stage for a two part series regarding reproductive surgery. The development, importance, and implications for reproductive surgery in the setting of assisted reproductive technology is discussed. Images and videos augment the authors' reflections of the past, present, and/or future. This series will not include a focus on uterine transplantation as this will be a future topic of its own., (Copyright © 2019 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

Research Question: Assisted reproduction laboratories record instrument performance periodically. No standardized guidelines have been produced for this activity despite mandatory auditing systems in several countries. This study of 36 laboratories in 12 different countries was conducted to assess differences and similarities between quality assurance programmes using an adaptable cloud-based quality-control app for instrument monitoring., Design: A total of 36 deidentified IVF laboratories that subscribed to the same quality-assurance application were studied. Data were evaluated based on instrument types allocated to 10 domains: incubators, gas tanks, warming surfaces, refrigerators and freezers, cryo-storage, environment, water purification, peripheral equipment, checklists and miscellaneous., Results: The incubator domain constituted the greatest proportion of parameters (35%), followed by surface warming instruments at 15%. Most incubator O 2 readings were monitored between 4.5 and 5.5%, and between 5.5 and 6.5% for CO 2 . The altitude of the laboratory was poorly correlated with the CO 2 setting. Incubator display and measured values of gases and temperature by built-in sensors vary considerably compared with third-party sensors. A quality-control diligence score or mean average data points was calculated for each laboratory. This score is independent of number of instruments or laboratory size. Higher scores were associated with laboratories in countries with government regulations and mandatory auditing systems., Conclusions: Major differences exist in instrument monitoring practices among laboratories. Although incubator monitoring is the largest domain, many other sensitive instruments are diligently monitored by most laboratories. International standardization and guidelines are needed., (Copyright © 2019 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Purpose: Screening for blood born viruses is routinely performed before fertility treatment in assisted reproduction technology (ART) clinics worldwide. It involves testing for human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis C virus (HCV), among others. Identifying patients with positive viral screening allows to refer them and their partners for appropriate counseling and treatment. The need for repeat viral screening and its required frequency have never been clearly established. In Israel, viral screening is mandatory and is repeated annually. Our aim was to determine the prevalence of HBV, HCV, and HIV seroconversion in patients with negative screening upon initiation of ART treatment., Methods: A retrospective analysis of viral screening tests of all fertility patients in a single assisted conception unit between 1997 and 2015., Results: During the study period, 2844 patients were treated at our clinic, out of whom 1945 patients met the inclusion criteria. The average length of treatment was 1.61 ± 0.81 years, during which female patients underwent screening tests 2.6 ± 0.9 times, and male patients 2.3 ± 1.2 times. No case of seroconversion to any of the three viruses was noted during the entire study period, resulting a seroconversion rate of 0%., Conclusions: Primary infection with HBV, HCV, or HIV is an extremely rare event among Israeli infertile patients, and the risk for seroconversion in this population is practically nil. Annual screening of both partners leads to substantial costs and appears to be futile. Our results question the current practice and support increasing the interval between screening tests in low-risk populations.

The Asiatic black bear (ABB; Ursus thibetanus ussuricus) is a globally endangered species, and measures to help increase their population are necessary. For the successful restoration of this species, artificial breeding as well as conservation translocation are considered important. The aims of the present study were to evaluate the feasibility and effectiveness of urethral catheterization (UC), which is effectively used in feline species, for semen collection from ABBs and establish the optimal protocol for semen collection via this technique. Seven clinically healthy, adult male ABBs (age, 6-13 years; weight, 130-180 kg) housed at the Species Restoration Technology Institute, Korea were included in this study. All study procedures were performed during the breeding season (June to August) over 3 consecutive years. Semen samples were collected once or three times from all bears by ultrasound-guided UC or electroejaculation (EE) under general anesthesia, and their characteristics, including sperm motility, were evaluated. The day of semen collection was defined as Day 0. The semen collected by the UC method was stored at 4 °C, and sperm motility was evaluated at the same time every day for 16 days. The successful collection rates for the UC and EE methods were 92.3% and 53.8%, respectively. The sperm concentration (4718.9 ± 1526.1 vs. 185.0 ± 34.2 × 10 6 /ml), total sperm count (1196.6 ± 955.5 vs. 100.9 ± 70.0 × 10 6 ), sperm motility score (4.39 ± 0.78 vs. 3.00 ± 1.73), viability (98.2 ± 2.3 vs. 82.7 ± 19.6), and the proportion of spermatozoa with intact acrosomes (92.2% ± 9.3% vs. 75.6% ± 10.6%) were higher with the UC method than with the EE method, whereas the proportion of spermatozoa with an abnormal morphology (23.1% ± 4.6% vs. 45.6% ± 19.5%) was lower with the former than with the latter. Over the course of cool storage, there was an overall decrease in the total motility, progressive motility, and viability, although viability was >50% until Day 10. These findings suggest that ultrasound-guided UC is a useful and feasible tool for the collection of high-quality semen from ABBs. The collected semen remains viable for up to 10 days, with high sperm motility maintained for up to 7 days, when stored at 4 °C., (Copyright © 2019 Elsevier Inc. All rights reserved.)

There is an increasing demand for genetically modified mice produced without crossing, for rapid phenotypic screening studies at the organismal level. For this purpose, generation of completely embryonic-stem-cell (ESC)-derived chimeric mice without crossing is now possible using a microinjection or aggregation method with 3i culture medium. However, the microinjection of ESCs into blastocyst, morula, or 8-cell-stage embryos requires a highly skilled operator. The aggregation method is an easier alternative, but the conventional aggregation protocol still requires special skills. To make the aggregation method easier and more precise, here we developed a micro-aggregation device. Unlike conventional 3-dimensional culture, which uses hanging-drop devices for aggregation, we fabricated a polystyrene funnel-like structure to smoothly drop ESCs into a small area (300-μm in diameter) at the bottom of the device. The bottom area was designed so that the surface tension of the liquid-air interface prevents the cells from falling. After aggregation, the cells can be recovered by simply exerting pressure on the liquid from the top. The microdevice can be set upon a regular 96-well plate, so it is compatible with multichannel pipette use or machine operation. Using the microdevice, we successfully obtained chimeric blastocysts, which when transplanted resulted in completely ESC-derived chimeric mice with high efficiency. By changing the number of ESCs in the aggregate, we found that the optimum number of co-cultured ESCs was around 90~120 per embryo. Under this condition, the efficiency of generating completely ESC-derived mice was the same or better than that of the injection method. These results indicated that our microdevice can be used to produce completely ESC-derived chimeric mice easily and with a high success rate, and thus represents a promising alternative to the conventional microinjection or aggregation method, especially for high-throughput, parallel experimental applications., Competing Interests: The authors have declared that no competing interests exist.

Various types of medical devices used in assisted reproductive technologies (ART) should be detected for their safety by strict biological assays. Mouse embryo assay(MEA)has been recognized as one of the most important and standardized methods with the threshold more than 80% of blastocyst formation rate (BR) after 96 h culture of fertilized eggs. The disadvantage using BR for embryonic quality control has been concerned as it is ubiquitously dependent of embryonic morphology and the detailed data including molecular and genetic information is obviously missing and incomplete. This leads to the urgent requirement for more sensitive and efficient assessments for the quality control of ART. This study evaluated the reliability of an immunofluorescent MEA by counting total cell and differential number of the cells in the inner cell mass (ICM) and trophectoderm (TE) in the blastocyst. This method improved the traditional MEA, provided a sensitive and powerful platform to assess embryonic developmental viability and should be suggested as a standard assay to be globally used for the quality control of medical devices and pre-clinical procedures in ART.

Background: Owing to the wide-ranging role nurses have in supporting patients undergoing fertility treatment, we recorded the learning/teaching expectations and experiences of nurses using a redesigned fertility pen injector., Methods: This was a multicentre, simulated-use study, using unbranded placebo-filled pens. Before teaching patients, nurses were given free choice to rank the importance of the device attributes and predict the level of patient anxiety. Nurses taught 2-5 patients how to prepare the device, inject the dose and complete an incomplete dose. They rated the teaching experience on a 5-point scale during a questionnaire interview., Results: Thirty nurses were enrolled across four countries. All nurses found the redesigned fertility pen injector easy to use and teach. 90% found the overall administration process easy to learn and teach. More than 80% (range 83%-100%) found each of the steps easy (score 4 or 5), and most found the steps easier to teach than expected (score 4 or 5; range 57%-90%). 97% would recommend the redesigned fertility pen injector to a colleague., Conclusions: Nurses rated the redesigned fertility pen injector easy to learn and use and easier to teach than expected. Most would recommend the device to a colleague.

Zona pellucida (ZP) manipulation, termed "assisted hatching" (AH), has been introduced in order to favor embryo hatching and ultimately improve assisted reproductive technology success but with poor proofs of safety and biological plausibility. We herein provide a systematic review of clinical outcomes following the application of different methods of ZP manipulation on fresh or frozen/thawed embryos at different developmental stages in different groups of patients. Out of the 69 papers that compared the clinical outcomes deriving from hatched versus non-hatched embryos, only 11 considered blastocysts while the rest referred to cleavage stage embryos. The ZP thinning of fresh embryos either by chemical or laser approach was shown to provide very limited benefit in terms of clinical outcomes. Better results were observed with procedures implying a higher degree of zona manipulation, including zona removal. Studies comparing the mechanical or chemical procedures to those laser-mediated consistently reported a superiority of the latter ones over the former. Literature is consistent for a benefit of ZP breaching in thawed blastocysts. This review provides the current knowledge on the AH procedure in order to improve its efficacy in the appropriate context. Embryologists might benefit from the approaches presented herein in order to improve Assisted Reproduction Technologies (ART) outcomes.

Kinetic (dynamic) vitrification is a promising trend in cryopreservation of biological materials because it allows avoiding the formation of lethal intracellular ice and minimizes harmful effects of highly toxic penetrating cryoprotectants. A uniform cooling protocol and the same instruments can be used for practically all types of cells. In modern technologies, the rate of cooling is essentially limited by the Leidenfrost effect. We describe a novel platform for kinetic vitrification of biological materials KrioBlast TM that realizes hyper-fast cooling and allows overcoming the Leidenfrost effect. This opens prospects for creation of a novel technology of cell cryopreservation for reproductive and regenerative medicine.

Assisted reproductive technology (ART) can benefit from the features of microfluidic technologies, such as the automation of time-consuming labor-intensive procedures, the possibility to mimic in vivo environments, and the miniaturization of the required equipment. To date, most of the proposed approaches are based on polydimethylsiloxane (PDMS) as platform substrate material due to its widespread use in academia, despite certain disadvantages, such as the elevated cost of mass production. Herein, we present a rapid fabrication process for a cyclic olefin copolymer (COC) monolithic microfluidic device combining hot embossing-using a low-temperature cofired ceramic (LTCC) master-and micromilling. The microfluidic device was suitable for trapping and maturation of bovine oocytes, which were further studied to determine their ability to be fertilized. Furthermore, another COC microfluidic device was fabricated to store sperm and assess its quality parameters over time. The study herein presented demonstrates a good biocompatibility of the COC when working with gametes, and it exhibits certain advantages, such as the nonabsorption of small molecules, gas impermeability, and low fabrication costs, all at the prototyping and mass production scale, thus taking a step further toward fully automated microfluidic devices in ART.

Measurement of oocyte and embryo biomechanical properties has recently emerged as an exciting new approach to obtain a quantitative, objective estimate of developmental potential. However, many traditional methods for probing cell mechanical properties are time consuming, labor intensive and require expensive equipment. Microfluidic technology is currently making its way into many aspects of assisted reproductive technologies (ART), and is particularly well suited to measure embryo biomechanics due to the potential for robust, automated single-cell analysis at a low cost. This review will highlight microfluidic approaches to measure oocyte and embryo mechanics along with their ability to predict developmental potential and find practical application in the clinic. Although these new devices must be extensively validated before they can be integrated into the existing clinical workflow, they could eventually be used to constantly monitor oocyte and embryo developmental progress and enable more optimal decision making in ART., (© The Author 2016. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Study Question: Do the embryo culture media and plastic materials used during assisted reproductive technology (ART) laboratory procedures expose embryos to bisphenol A (BPA)?, Summary Answer: BPA was not detected in embryo culture media or protein supplements at concentrations above those encountered in normal patient serum and follicular fluids., What Is Known Already: BPA is strongly suspected of altering the epigenome during mammalian development. Medical devices have been shown to be a source of BPA exposure in adult and neonatal intensive care units., Study Design, Size, Duration: An analytical study of ART culture media and plastic labware products was performed under conditions close to routine practice and if BPA was detected, tests were carried out under more stringent conditions., Participants/materials, Setting, Methods: Two single-step embryo culture media, two sequential media and three different protein supplements [a purified human serum albumin (HSA), a synthetic serum substitute, and a recombinant HSA] were tested for BPA. Thirty-three different plastic consumables, used from oocyte collection through to embryo transfer, were tested for their ability to leach BPA into their surrounding environment.BPA concentrations were measured according to a previously described liquid chromatography/mass spectrometry method. This method is linear over the calibration range from 0.5 to 100 ng/ml using a linear model weighted by 1/X² and validated in terms of selectivity, linearity, repeatability, reproducibility and limit of quantification (0.5 ng/ml)., Main Results and the Role of Chance: Neither the culture media nor the protein supplements were shown to contain detectable levels of BPA. None of the plastic materials leached BPA into the surrounding medium at levels higher than the upper limit detected previously in serum and follicular fluids in women (about 2 ng/ml). However, the plastic of the three tested strippers used for oocyte denudation/embryo handling did contain BPA. Two of these strippers are made with polycarbonate, a plastic whose synthesis is known to require BPA., Limitations, Reasons for Caution: This study is limited to the ART media and materials tested here and using a BPA assay with a limit of quantification at 0.5 ng/ml. A minimum volume was required for testing, and one type of plastic labware could not be tested in conditions identical to those in routine use., Wider Implications of the Findings: Although we demonstrated that some plastic materials used in ART contain BPA, under routine conditions none appear capable of leaching BPA at levels higher than those from maternal internal exposure. However, BPA is strongly suspected of altering the epigenome. Since important epigenetic modifications occur in the early embryonic stage, it is questionable whether plastics that contain BPA, polycarbonate in particular, should be used in the manufacture of plastic consumables for ART procedures., Study Funding/competing Interests: This work was supported by a grant from the Agence de Biomédecine (AOR 2012) and by a grant from the French Ministry of Health (Clinical Research Hospital Program 2012; no.12-018-0560). The authors declared no competing interest., (© The Author 2016. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

This postcoital sperm assessment study was performed over a 10 month time period (November 2014-August 2015). Fifteen couples enrolled in the study. The study was a non-blinded, non-randomized, single-center comparison study comparing The Stork® OTC (Rinovum Women's Health, Monroeville, PA) to natural intercourse (NI), using the subjects as their own control/baseline. This was an efficacy study designed to compare the number of sperm in the cervical mucus following the use of The Stork OTC conception aid with the number of sperm in the cervical mucus following natural intercourse. Subjects used both The Stork OTC conception system and the natural intercourse method to evaluate concentrations of sperm in the cervical mucus. Post-coital test (PCT) data was collected demonstrating higher concentrations of sperm within the cervical mucus with The Stork OTC conception system versus natural intercourse for 85% of test subjects in this study. Of the 15 couples enrolled in the study, 2 were lost to follow-up. Mean age for male subjects was 31.7 +/ 5.4 years of age and mean age for female subjects was 29.7+/- 5.4. The average sperm score value of the 85% of test subjects with higher sperm concentrations from The Stork OTC was 3.23 times the score value of sperm concentration compared to natural intercourse. The remaining 15% of test subjects showed no change in sperm score value between The Stork OTC and natural intercourse.

15% of cancers affecting women are of gynaecologic origin, thereby most of them affect fertility of the women who are in reproductive age indirectly by their treatment protocols including chemotherapy, radiotherapy etc. Chances of getting pregnant is the crucial point of concern in these patients as pathological state of these women potentially limits chances of giving birth to new born. However, current advances in our understanding of these diseases, along with improved multimodality treatments, allow for consideration of fertility options. So, the present review shall enlighten technological advancements available including robotic surgical procedures, fertility preservation innovations and other latest developments in the area for improvising fertility options in gynaecological cancer patients.

We previously demonstrated that tauroursodeoxycholic acid (TUDCA) improved the developmental competence of mouse embryos by attenuating endoplasmic reticulum (ER) stress-induced apoptosis during preimplantation development. Here, we present a follow-up study examining whether TUDCA enhances the implantation and live-birth rate of mouse embryos. Mouse 2-cell embryos were collected by oviduct flushing and cultured in the presence or absence of 50 μM TUDCA. After culture (52 h), blastocysts were transferred to 2.5-day pseudopregnant foster mothers. It was found that the rates of pregnancy and implantation as well as the number of live pups per surrogate mouse were significantly higher in the TUDCA-treated group compared to the control group, but there was no significant difference in the mean weights of the pups or placentae. Thus, we report for the first time that TUDCA supplementation of the embryo culture medium increased the implantation and livebirth rates of transferred mouse embryos., (Copyright © 2015 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.)

The Food and Drug Administration (FDA) is classifying the Assisted Reproduction Embryo Image Assessment System into class II (special controls). The special controls that will apply to the device are identified in this order, and will be part of the codified language for the Assisted Reproduction Embryo Image Assessment System classification. The Agency is classifying the device into class II (special controls) in order to provide a reasonable assurance of safety and effectiveness of the device.

Purpose: Vitrification significantly improves the rates of blastocyst survival and clinical pregnancy following frozen embryo transfer (FET). However, ice crystal formation during the freezing process reduces the blastocyst survival rate. Artificial shrinkage (AS) prior to blastocyst vitrification decreases the formation of ice crystals, increasing the blastocyst survival rate. The aim of this study was to identify an efficient AS method to improve blastocyst survival rates following vitrification., Method: Use of the 29-gauge needle AS and Laser pulse AS methods prior to vitrification was compared in terms of the impacts on the rates of blastocyst survival in FET cycles, blastocyst hatching, clinical pregnancy after transfer, embryo implantation, abortion, gestational duration and birth weight., Result: In total, 438 blastocysts in 219 cycles were thawed, resulting in survival of 407 (92.9 %). Of these, 213 cycles were transferred, resulting in 129 clinical pregnancies (60.6 %) and 140 successful births. There were no differences between the two methods in the rates of blastocyst survival, clinical pregnancy, embryo implantation and abortion. However, the 29-gauge needle AS group was associated with a significantly lower blastocyst hatching rate (83.6 % vs. 91.2 %), shorter average gestational duration (37.36 ± 2.34 vs. 38.06 ± 1.76), and higher premature birth rate (40.00 % vs. 21.15 %) compared with Laser pulse AS group., Conclusion: No significant differences in the effectiveness of the two methods applied prior to blastocyst vitrification were observed before birth, while after birth, a significantly improved clinical outcome was obtained with laser pulse AS indicating that this is a more effective pre-processing method for blastocyst vitrification.

The objective of this study was to evaluate the efficacy of a novel polydimethylsiloxane (PDMS) well-insert system for oocyte in vitro maturation (IVM) and in vitro embryo culture (IVC) in pigs. The PDMS well inserts, consisting of multiple microwells with connecting microchannels, resulted in equivalent blastocyst development compared with standard microdrop culture for IVC. These PDMS well inserts were then evaluated for IVM or IVC in a rocking versus static environment. The rocking environment during both oocyte IVM and embryo culture had detrimental effects on oocyte and embryo development compared with a static environment. Importantly, blastocyst development of oocytes and embryos cultured in the PDMS well inserts in the static environment was equivalent to that of standard microdrops. Further analysis of transcript abundance in blastocysts produced from these different environments revealed that the PDMS well-insert system may produce more viable embryos. In conclusion, this PDMS well-insert system can successfully mature oocytes and culture embryos in an individually-identifiable manner without compromising, and perhaps enhancing, developmental potential.

Study Question: Is post-implantation embryonic development after blastocyst transfer affected by exposure to different assisted reproduction technology (ART) culture media?, Summary Answer: Fetal development and placental histology of ART embryos cultured in vitro in different ART media was not impaired compared with embryos grown in vivo., What Is Known Already: The application of different in vitro culture (IVC) media for human ART has an effect on birthweight of newborns. In the mouse model, differences in blastocyst formation were reported after culture in different ART media. Moreover, abnormalities in the liver and heart have been detected as a result of suboptimal IVC conditions., Study Design, Size, Duration: Fertilized oocytes from inbred and outbred breeding schemes were retrieved and either immediately transferred to foster mothers or incubated in control or human ART culture media up to the blastocyst stage prior to transfer. Placental and fetal anatomy and particularly bone development were evaluated., Participants/materials, Setting, Methods: B6C3F1 female mice were used as oocyte donors after ovulation induction. C57Bl/6 and CD1 males were used for mating and CD1 females as foster mothers for embryo transfer. Fertilized oocytes were recovered from mated females and incubated in sequential human ART media (ISM1/ISM2 and HTF/Multiblast), in control media [KSOM(aa) and Whitten's medium] or grown in utero without IVC (zygote control). As in vivo, control B6C3F1 females were superovulated and left untreated. Fetuses and placentae were isolated by Caesarean section and analysed at 18.5 days post-coitum (dpc) for placenta composition and at 15.5 dpc for body weight, crown-rump length (CRL), fetal organ development, morphological development, total bone length and extent of bone ossification., Main Results and the Role of Chance: No major differences in the number of implantation sites or in histological appearance of the placentae were detected. CRL of KSOM(aa) fetuses was higher compared with zygote control and Whitten's medium. Histological analysis of tissue sections revealed no gross morphological differences compared with the in vitro groups or in vivo controls. Furthermore, no changes in skeletal development and degree of ossification were observed. However, fibula and tibia of ISM1/ISM2 fetuses were longer than the respective ones from in vivo fetuses., Limitations, Reasons for Caution: Findings in the mouse embryo and fetus may not be fully transferable to humans. In addition to skeletal development and placentation, there may be other parameters, e.g. on the molecular level which respond to IVC in ART media. Some comparisons have limited statistical power., Wider Implications of the Findings: Our data suggest that once implantation is achieved, subsequent post-implantation development unfolds normally, resulting in healthy fetuses. With mouse models, we gather information for the safety of human ART culture media. Our mouse study is reassuring for the safety of ART conditions on human embryonic development, given the lack of bold detrimental effects observed in the mouse model., Study Funding/competing Interest(s): This work was supported by the Deutsche Forschungsgemeinschaft (BO 2540/4-1 and SCHL 394/9-1) and by the Nederlandse Organisatie voor Wetenschappelijk Onderzoek (S.L.G.); Bilateral grant NWO-DFG 63-258. None of the authors has any conflict of interest to declare., Trial Registration Number: Not applicable.

Purpose: To determine if there is a temperature variation within and between incubators., Methods: This prospective, experimental trial with external controls was performed at an Assisted Reproductive Technology laboratory in a tertiary-care, university hospital. Temperature values were taken at various locations within and between incubators., Results: Even though they were both set to 37.0 °C, the same make and model incubators had significantly different internal temperatures. Temperatures differed significantly among top, middle and bottom shelves and between fronts and backs of shelves., Conclusion(s): We found temperatures differed within and between our front-loading incubators. Thus, laboratory personnel should evaluate their incubators for temperature variations within and between incubators and, if temperatures differ significantly, develop a plan to deal with discrepancies.

Objective: We evaluated the ease of use of a pen injector for follitropin α (recombinant human follicle-stimulating hormone [r-hFSH]) during assisted reproduction technologies (ARTs) in Egypt., Methods: One hundred women undergoing ART completed a questionnaire in a non-interventional, observational study. The primary endpoint was patients' rating of the comfort associated with the injector. The main limitations of the study were the design and lack of knowledge regarding any impact of failure of ART on perceptions of treatment for a minority of patients., Results: Patients rated the follitropin α pen injector as 'very comfortable' (61%), 'comfortable' (29%), or 'somewhat comfortable' (10%). Understanding instructions and using it were 'very easy' or 'easy' for 97-99%; 94% reported 'no' or 'minimal' difficulty with injections, 83% were 'very confident' about altering doses, 77% reported no interference with normal daily activities and 94% reported 'no' or 'minimal' stress using the device. Women with previous experience of ART rated the device as more practical than their previous injection system. Overall, 96% were 'very satisfied' or 'satisfied' with the device and 99% would recommend its use to others. Pregnancy rates were consistent with previous clinical experience. Injection site reactions occurred in 10% (all of mild severity except one moderate event)., Conclusions: Positive perceptions of the follitropin α pen injector identify this device as suitable for use for Middle Eastern women undergoing ART.

We performed a systematic review and meta-analysis to examine whether a difficult embryo transfer or the presence of blood on the transfer catheter affects assisted reproduction outcomes. We searched the following databases: Cochrane Central Register of Controlled Trials (CENTRAL), Database of Abstracts of Reviews of Effects (DARE), MEDLINE, Embase, PsycINFO, Cumulative Index to Nursing and Allied Health Literature (CINAHL), and Literatura Latino-Americana e do Caribe em Ciências da Saúde (LILACS). We aimed to determine the risk ratio (RR) associated with difficult embryo transfer or the presence of blood on the transfer catheter for the following outcomes: live birth, clinical pregnancy, and miscarriage. We identified 3066 papers, of which 194 were reviewed and nine were included. The outcome of live birth was not reported in any of the included studies and the effect on miscarriage was too imprecise for any conclusions. Pooled analysis of five studies demonstrated lower clinical pregnancy rates following a non-easy embryo transfer (RR=0.75; 95% CI=0.66-0.86). This included three studies showing subjectively difficult transfers reducing clinical pregnancies (RR=0.67; 95% CI=0.51-0.87) and two studies in which the need for additional manoeuvers reduced clinical pregnancies (RR=0.78; 95% CI=0.67-0.91). The presence of blood on the transfer catheter did not affect clinical pregnancy rates (RR=0.96; 95% CI=0.82-1.14) in five studies. We concluded that low quality evidence suggests that a difficult embryo transfer but not a bloody catheter reduces the chance of achieving a clinical pregnancy. More good quality studies are needed to evaluate the effect of difficult embryo transfer and the presence of blood on the catheter on the main outcomes of assisted reproduction., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

Background: Gonadotrophins are used routinely for follicular stimulation during ovarian induction and assisted reproduction techniques. Developments in recombinant follicle-stimulating hormone preparations and their injection devices have improved patient quality of life by enabling patients to self-administer treatment at home. The objective of this study was to investigate patient experiences of learning to use and overall satisfaction with the follitropin-alpha (Gonal-f) filled-by-mass (FbM) prefilled pen., Methods: This questionnaire-based survey study was conducted in 23 fertility centres in Japan over a period of 14 months. Patients who were receiving fertility treatment with the follitropin-alpha (FbM) prefilled pen were asked to complete a questionnaire to assess their satisfaction, ease of learning and use, and injection site pain following treatment., Results: A total of 663 women participated in the study. The majority of patients found the instructions for administering follitropin-alpha with the prefilled pen easy to understand (83.0%; n = 546/658) and patients found that a hands-on demonstration by a nurse or doctor was the most useful tool for learning to use the follitropin-alpha (FbM) prefilled pen (80.0%; n = 497/621). Forty-eight percent (n = 318) of patients in the study had previous experience with different types of fertility medications and the majority of these patients found the follitropin-alpha (FbM) prefilled pen easier to use (75.1%; n = 232/309) and less painful (89.0%; n = 347/390) than their previous medication. The majority (80.2%; n = 521/650) of patients reported overall satisfaction with the follitropin-alpha (FbM) prefilled pen., Conclusions: The follitropin-alpha (FbM) prefilled pen is an easy-to-use injection device according to this questionnaire-based survey. Patients who had experience of different types of fertility medication preferred the follitropin-alpha (FbM) prefilled pen to other injection devices.

The objective of this study is to present results for measurement of estradiol (E(2)) in serum and control materials with a new automated assay (eE(2)) that could be used in random continuous access mode on the Siemens laboratories Advia-Centaur immunoassay analyzer. We evaluated the imprecision (within and between run), the linearity, the limit of detection, and the functional sensitivity. We have tested the assay for monitoring ovulation stimulation for in vitro fertilization and embryo transfer (IVF-ET) throughout the most frequent protocol utilized in our IVF centre (n = 247). Results are compared to those obtained by the E(2)6 assay, one other assay on the Advia-Centaur immunoassay analyzer (Siemens), routinely used in our hormonology department. The obtained results show that the new assay presents better analytical performances (precision, linearity, limits of detection and sensibility) than the previous technique.

Objective: This retrospective case-control study was undertaken to assess the impact of a malleable stylet application on embryo transfer (ET) outcome., Methods: The Wallace malleable stylet was used when the soft inner catheter could not negotiate the internal os during ultrasound-guided ET. Patients having an age ≤37 years, with three embryos replaced were included in the analysis. Implantation rate, clinical pregnancy rate and live birth rate were the main outcome measures., Results: The Wallace malleable stylet was applied in 164 (29.6%) of the 553 ETs evaluated. Implantation rate, clinical pregnancy rate and live birth rate showed a 5.6% (19.4% vs. 13.8%), a 10.8% (41.9% vs. 31.1%) and a 9.9% (37.3% vs. 27.4%) absolute difference, respectively, in favor of ETs not requiring the stylet. These differences were statistically significant (P < 0.05)., Conclusions: ETs requiring stylet are associated with lower implantation, clinical pregnancy and live birth rates.

The growth of mammalian embryos in the laboratory has progressed nicely over the last 60 to 70 years. This has been important for basic studies of preimplantation embryo regulation of growth, development, and differentiation. It has also been critical for the establishment of assisted reproductive technologies in the clinical setting to treat infertility. During the first 40 to 50 years, the primary focus was on defining soluble media components, which fortunately has yielded improved success in the culture of embryos. More recently, attention has shifted to other aspects of culture beyond media composition. In this review we discuss recent advances in preimplantation embryo culture that deal with modifying the insoluble culture environment, producing culture environments that are dynamic and not static, systems that allow more precise gas phase and/or media pH regulation/monitoring, and platforms that integrate with culture systems to allow analysis of embryos and thus assist in selection of the embryos with the greatest implantation potential., (Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.)

The preponderance of research toward improving embryo development in vitro has focused on manipulation of the chemical soluble environment, including altering basic salt composition, energy substrate concentration, amino acid makeup, and the effect of various growth factors or addition or subtraction of other supplements. In contrast, relatively little work has been done examining the physical requirements of preimplantation embryos and the role culture platforms or devices can play in influencing embryo development within the laboratory. The goal of this review is not to reevaluate the soluble composition of past and current embryo culture media, but rather to consider how other controlled and precise factors such as time, space, mechanical interactions, gradient diffusions, cell movement, and surface interactions might influence embryo development. Novel culture platforms are being developed as a result of interdisciplinary collaborations between biologists and biomedical, material, chemical, and mechanical engineers. These approaches are looking beyond the soluble media composition and examining issues such as media volume and embryo spacing. Furthermore, methods that permit precise and regulated dynamic embryo culture with fluid flow and embryo movement are now available, and novel culture surfaces are being developed and tested. While several factors remain to be investigated to optimize the efficiency of embryo production, manipulation of the embryo culture microenvironment through novel devices and platforms may offer a pathway toward improving embryo development within the laboratory of the future.

In vitro fertilisation (IVF) and related technologies are arguably the most challenging of all cell culture applications. The starting material is a single cell from which one aims to produce an embryo capable of establishing a pregnancy eventually leading to a live birth. Laboratory processing during IVF treatment requires open manipulations of gametes and embryos, which typically involves exposure to ambient conditions. To reduce the risk of cellular stress, we have developed a totally enclosed system of interlinked isolator-based workstations designed to maintain oocytes and embryos in a physiological environment throughout the IVF process. Comparison of clinical and laboratory data before and after the introduction of the new system revealed that significantly more embryos developed to the blastocyst stage in the enclosed isolator-based system compared with conventional open-fronted laminar flow hoods. Moreover, blastocysts produced in the isolator-based system contained significantly more cells and their development was accelerated. Consistent with this, the introduction of the enclosed system was accompanied by a significant increase in the clinical pregnancy rate and in the proportion of embryos implanting following transfer to the uterus. The data indicate that protection from ambient conditions promotes improved development of human embryos. Importantly, we found that it was entirely feasible to conduct all IVF-related procedures in the isolator-based workstations.

Purpose of Review: Since its inception in early 2000, robotic assistance with urologic procedures continues to expand. The magnification, three-dimensional visualization, and surgical control offered by the latest daVinci Si-HD system has led to its integration into microsurgical procedures for male infertility. The addition of robotic assistance may allow an improvement in outcomes similar to when the operating microscope was introduced in microsurgery. Though the use of robotics in microsurgery is still in its early phases, initial findings are encouraging., Recent Findings: This review covers robotic microsurgical procedures and tools for infertility and chronic orchialgia/testicular pain such as vasovasostomy, vasoepididymostomy, varicocelectomy, testicular sperm extraction and targeted denervation of the spermatic cord. Preliminary human clinical studies appear to show improved operative efficiency and comparable outcomes. The use of robotic assistance during robotic microsurgical vasovasostomy appears to decrease operative duration and improve the rate of return of postoperative sperm counts compared to the pure microsurgical technique., Summary: Long-term prospective controlled trials are necessary to assess the true benefit for robotic-assisted microsurgery. The preliminary findings are promising, but further evaluation is warranted.

This article explores the construction of meaning regarding assisted reproductive technology by legal framers, medical practitioners and would-be parents, through the concept of ecology of knowledge. It is argued that these inter-relationships between experts and lay people can be understood in terms of the formation of a social structure of ecology of knowledge, which depends on local and emotional knowledge co-produced by medical doctors, jurists and lay people in dynamic ways without compromising the autonomy of medical, legal and lay knowledge and skills. The assessment of the benefits and risks of assisted reproductive technology partially represents negotiations of knowledge between these social and professional groups, aiming to reproduce existing relations and practices, particularly the social power of medicine and technology, the dominant perceptions about women's and men's bodies and the geneticisation of genealogy. These negotiations of knowledge generate new rights, new social actors, new scientific fields and new ways of thinking and talking about individual and institutional responsibilities. Ecology of knowledge comes imbued with hope, trust, power, credibility of institutions and moralisation whereby some citizens' rights may be weakened., (© 2011 The Authors. Sociology of Health & Illness © 2011 Foundation for the Sociology of Health & Illness/Blackwell Publishing Ltd.)

During the past decades, improvements in culture of preimplantation embryos have contributed substantially in the success of human assisted reproductive techniques. However, most efforts were focused on optimization of media and gas components, while the established physical conditions and applied devices have remained essentially unchanged. Very recently, however, intensive research has been started to provide a more appropriate environment for the embryos and to replace the rather primitive and inappropriate devices with more sophisticated and practical instruments. Success has been reported with simple or sophisticated tools (microwells or microchannels) that allow accumulation of autocrine factors and establishment of a proper microenvironment for embryos cultured individually or in groups. The microchannel system may also offer certain level of automation and increased standardization of culture parameters. Continuous monitoring of individual embryos by optical or biochemical methods may help to determine the optimal day of transfer, and selection of the embryo with highest developmental competence for transfer. This advancement may eventually lead to adjustment of the culture environment to each individual embryo according to its actual needs. Connection of these techniques to additional radical approaches as automated ICSI or an ultimate assisted hatching with full removal of the zona pellucida after or even before fertilization may result in devices with high reliability and consistency, to increase the overall efficiency and decrease the work-intensity, and to eliminate the existing technological gap between laboratory embryology work and most other fields of biomedical sciences., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The meiotic spindle in the oocyte is composed of microtubules and plays an important role during chromosome alignment and separation at meiosis. Polarized light microscopy (PLM) could be useful for a non-invasive evaluation of the meiotic spindle and may allow removal of nuclear structures without fluorochrome staining and ultraviolet exposure. In this study, PLM was used to assess its potential application in porcine reproductive technologies. The objectives of the present study were to assess the efficiency of PLM to detect microtubule-polymerized protein in in vitro-matured porcine oocytes; to examine its effects on the oocyte developmental competence; to select oocytes based on the presence of the meiotic spindle detected by PLM; and to assess the efficiency oocyte enucleation assisted with PLM. In the first experiment, the presence of microtubule-polymerized protein was assessed and confirmed in oocytes (n = 117) by immunostaining and chromatin detection. In the second experiment, oocytes (n = 160) were exposed or not (controls) to PLM for 10 minutes, and then parthenogenetically activated and cultured in vitro. In the third experiment, development competence of oocytes with a positive or negative signal to PLM was analyzed after in vitro fertilization. Finally, oocytes (n = 54) were enucleated using PLM as a tool to remove the meiotic spindle. A positive PLM signal was detected in 98.2 % of the oocytes, which strongly correlated (r = 1; p < 0.0001) with the presence of microtubule-polymerized protein as confirmed by immunostaining. Oocytes exposed to PLM did not differ significantly from controls on cleavage, total blastocyst, expanded blastocyst rates and total cell numbers. The percentage of oocytes at the MII stage and blastocyst formation rate in the negative PLM group significantly differed from control and PLM positive groups. Overall efficiency of spindle removal using the PLM-Oosight system was 92.6%. These results suggest that polarized light microscopy is an efficient system to detect microtubule-polymerized protein in in vitro-matured porcine oocytes and does not exert detrimental effects on porcine oocyte developmental competence. Selecting oocytes by the presence of a PLM signal provides limited improvement on IVF results. Finally, PLM appears as an efficient method to enucleate porcine oocytes., (Copyright © 2011 Elsevier Inc. All rights reserved.)

The aim of this report is to describe our experience and results with implementation of a cut standard straw technique for vitrification and warming of day 3 cleavage-stage human embryos. Detailed description of the method and results of 63 frozen embryo transfers performed with this technology are discussed, and it is concluded that this method provides a reliable, inexpensive, and effective option of embryo vitrification at a cleaved stage., (Copyright © 2011 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

The Great Eastern Japan Earthquake of 2011 and consequent tsunami affected many IVF clinics in northeastern Japan. Twelve clinics lost at least one embryo, but all the frozen embryos stored in liquid nitrogen tanks were safe. Emergency power supply is the first priority issue in an unpredictable natural disaster., (Copyright © 2011 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

Objective: To evaluate the safety of a new ultravitrification closed device., Design: Ultravitrification research., Setting: Private assisted reproduction center., Animal(s): Microorganisms (bacteria)., Intervention(s): A styrofoam container holding 1,000 mL of liquid nitrogen (LN2) was contaminated with Pseudomonas aeruginosa and Escherichia coli. Forty closed devices (Ultravit) and 20 open devices (Cryotop) loading approximately 0.5 μL of antibiotic-free medium were plunged into this contaminated LN2 for 5-10 seconds and then inoculated into selective culture dishes. Colony-forming units were analyzed and counted after an overnight incubation at 37°C., Main Outcome Measure(s): Detection of micro-organisms in different devices after incubation., Result(s): There was no contamination in any of the closed devices, whereas in 45% of open devices these bacteria were present., Conclusion(s): With this study we demonstrated, in an experimental model using contaminated LN2, that this new closed device is a safe system that does not allow cell contact with LN2, avoiding cell contamination., (Copyright © 2011 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

Preservation of female genetics is currently done primarily by means of oocyte and embryo cryopreservation. The field has seen much progress during its four-decade history, progress driven predominantly by research in humans, cows, and mice. Two basic cryopreservation techniques rule the field--controlled-rate freezing, the first to be developed, and vitrification, which, in recent years, has gained a foothold. While much progress has been achieved in human medicine, the cattle industry, and in laboratory animals, this is far from being the case for most other mammals and even less so for other vertebrates. The major strides and obstacles in human and other vertebrate oocyte and embryo cryopreservation will be reviewed here.

In vitro ovarian follicle culture is a new frontier in assisted reproductive technology with tremendous potential, especially for fertility preservation. Folliculogenesis within the ovary is a complex process requiring interaction between somatic cell components and the oocyte. Conventional two-dimensional culture on tissue culture substrata impedes spherical growth and preservation of the spatial arrangements between oocyte and surrounding granulosa cells. Granulosa cell attachment and migration can leave the oocyte naked and unable to complete the maturation process. Recognition of the importance of spatial arrangements between cells has spurred research in to three-dimensional culture system. Such systems may be vital when dealing with human primordial follicles that may require as long as three months in culture. In the present work we review pertinent aspects of in vitro follicle maturation, with an emphasis on tissue-engineering solutions for maintaining the follicular unit during the culture interval. We focus primarily on presenting the various 3-dimensional culture systems that have been applied for in vitro maturation of follicle:oocyte complexes. We also try to present an overview of outcomes with various biomaterials and animal models and also the limitations of the existing systems.

Background: Self-administration of recombinant human follicle-stimulating hormone (r-hFSH) can be performed using injection pen devices by women undergoing assisted reproductive technology procedures. The objective of this study was to explore the use of the prefilled follitropin alfa pen in routine assisted reproductive technology procedures in Germany., Methods: This prospective, observational study was conducted across 43 German IVF centres over a period of 1.75 years. Patients who had used the prefilled follitropin alfa pen in the current or a previous cycle of controlled ovarian stimulation completed a questionnaire to assess their opinions of the device., Results: A total of 5328 patients were included in the study. Of these, 2888 reported that they had previous experience of daily FSH injections. Significantly more patients reported that less training was required to use the prefilled follitropin alfa pen than a syringe and lyophilized powder (1997/3081 [64.8%]; p < 0.001 'less' versus 'more' training). Significantly more patients rated the prefilled follitropin alfa pen as easier in terms of use (2321/3206, 72.4%; p < 0.001 'much more easy' versus 'less easy') and daily injection (2384/3262, 73.1%; p < 0.001 'much more easy' versus 'less easy') than existing injection methods. Approximately one third of respondents rated the prefilled follitropin alfa pen as easier to use than the follitropin beta pen with reloadable cartridges. The majority (3378/4024, 83.9%) of patients had a general preference for the prefilled follitropin alfa pen over other injection methods., Conclusions: In this questionnaire-based survey, routine use of the prefilled follitropin alfa pen was well accepted and associated with favourable patient perceptions. Users of the pen found it easier to initially learn how to use, and subsequently use, than other injection methods. In general, the prefilled follitropin alfa pen was the preferred method for self-administration of gonadotrophins. Together with previous findings, the results here indicate a high level of patient satisfaction among users of the prefilled follitropin alfa pen for daily self-administration of r-hFSH.

Objective: To identify common clinical and laboratory practices among consistently high-performing IVF programs., Design: Questionnaire study of selected IVF programs., Setting: Academic and private practice IVF programs., Patient(s): Ten of 12 programs identified as having consistently high singleton delivery rates per cycle., Intervention(s): None., Main Outcome Measure(s): Common clinical practices., Result(s): Common clinical practices identified among these programs included testing all patients for ovarian reserve, endometrial defects, and hydrosalpinges; use of a mixed LH and FSH stimulation protocol with step-down dosing; and use of ultrasound guidance for ET. Common laboratory practices included selective use of intracytoplasmic sperm injection, group culture of embryos in microdrops, and use of blastocyst ET in selected cases. Common laboratory features included good air quality using filtration and heated stages for oocyte and embryo work., Conclusion(s): Although a number of factors were identified in this best-practices questionnaire, programs often differed in many aspects of care. However, high-performing programs cited experience of physicians, embryologists, and staff members as well as consistency of approach, attention to detail, and good communication as being vital to excellent outcomes., (Copyright (c) 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

In 2006 the Dutch Society for Clinical Embryologists (KLEM) approached the Netherlands Standardisation Institute (NEN) for advice regarding the lack of safety and quality specifications for medical devices used in assisted reproductive technology. A project plan was drafted in accordance with NEN-standardized methods for the development of norms and Dutch technical specifications (NTA) and a working group was launched consisting of all interested parties. A framework was then set up to develop an NTA that focused on the safety of gametes, embryos and the unborn offspring. The three main parts of the NTA describe the classification of medical devices, the requirements for new and existing devices and testing methods. The content of the NTA can be considered mainly as a consensus of the participants in the working group from both industry and clinical embryologists (KLEM). A final draft was sent to representatives from or allied to the government and to a notified body, and NTA 8070, entitled Devices for Assisted Reproductive Technologies (ART), was issued in March 2008., (2010 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.)