Assisted Reproductive Procedures Research Articles

The activation of the chymotrypsin-like activity of the proteasome is regulated by soluble adenyl cyclase/cAMP/protein kinase A pathway and required for human sperm capacitation.

One of the first events of mammalian sperm capacitation is the activation of the soluble adenyl cyclase/cAMP/protein kinase A (SACY/cAMP/PKA) pathway. Here, we evaluated whether the increase in PKA activity at the onset of human sperm capacitation is responsible for the activation of the sperm proteasome and whether this activation is required for capacitation progress. Viable human sperm were incubated with inhibitors of the SACY/cAMP/PKA pathway. The chymotrypsin-like activity of the sperm proteasome was evaluated using a fluorogenic substrate. Sperm capacitation status was evaluated using the chlortetracycline assay and tyrosine phosphorylation. To determine whether proteasomal subunits were phosphorylated by PKA, the proteasome was immunoprecipitated and tested on a western blot using an antibody against phosphorylated PKA substrates. Immunofluorescence microscopy analysis and co-immunoprecipitation (IPP) were used to investigate an association between the catalytic subunit alpha of PKA (PKA-Cα) and the proteasome. The chymotrypsin-like activity of the sperm proteasome significantly increased after 5min of capacitation (P< 0.001) and remained high for the remaining incubation time. Treatment with H89, KT5720 or KH7 significantly decreased the chymotrypsin-like activity of the proteasome (P< 0.001). IPP experiments indicated that PKA inhibition significantly modified phosphorylation of proteasome subunits. In addition, PKA-Cα colocalized with the proteasome in the equatorial segment and in the connecting piece, and co-immunoprecipitated with the proteasome. This is the first demonstration of sperm proteasome activity being directly regulated by SACY/PKA-Cα. This novel discovery extends our current knowledge of sperm physiology and may be used to manage sperm capacitation during assisted reproductive technology procedures.

Jsou všichni lidé osoby, a jestliže ano, proč?: Náčrt SKIP argumentů a argumentace Roberta Spaemanna

The question of whether all people (i.e., members of homo sapiens) are persons returns whenever some people are, in practice, excluded from the community of persons and when at the same time it is claimed that people have human dignity and basic human rights only as much as they are persons. This question is still topical, for example, for the moral evaluation of some assisted reproduction procedures and embryo­‑destructive research. The thesis that all people are persons is currently defended and justified by the SKIP-arguments. The article outlines the SKIP-arguments in propedeutic form and the argumentation of Robert Spaemann parallel to these arguments, who defends the thesis in an thoughtful way in his book Persons: The Difference between ’Someone‘ and ’Something‘. In the end, the reader will also learn about the meta-argument of tutiorism through which the advocates of this thesis usually strengthen the SKIP arguments.

Responsiveness of the cheetah (Acinonyx jubatus) ovary to exogenous gonadotropins after preemptive oral progestin treatment

Control of ovarian function in cheetahs is sub-optimal, which currently limits the integration of assisted reproductive techniques into the genetic management of that endangered species. The objective of this study was to determine the effect of preemptive progestin treatment on the quality of ovarian responses after exogenous gonadotropin stimulation in cheetahs. Adult females received either 1) 200 IU equine chorionic gonadotropin (eCG) followed with 3,000 IU porcine luteinizing hormone (pLH) (intramuscular route) (n = 5; control group) or 2) similar eCG/pLH administration preceded by a 7-day treatment with oral progestin (0.1 mg/kg altrenogest; ALT group; n = 7). At 42 h post-pLH administration, a series of metrics was assessed via laparoscopy (number of follicles ≥ 2 mm, number of corpora lutea, oviduct and uterine cornua diameter and overall vascularization). Concentrations of fecal estradiol, progesterone and glucocorticoid metabolites (FEM, FPM, and FGM, respectively) were measured by enzyme immunoassay for 3 wk before ALT treatment (Period 1), 7 d during ovarian suppression period (Period 2), throughout eCG/LH treatment and laparoscopy (Period 3), and 6 wk following laparoscopy (Period 4). Overall, nine out of 12 cheetahs (4/5 in control and 5/7 ALT group) had freshly-formed corpora lutea at the time of laparoscopy. Mean follicle and corpora lutea numbers in the control versus ALT group were not different (P > 0.05). Overall measurements and vascularization scores also did not differ (P > 0.05) among groups. FEM average concentrations increased (P ≤ 0.05) in response to eCG for the ALT-treated females between Periods 2 and 3 and were sustained during Period 4. However, FEM average concentrations did not vary (P > 0.05) for control females throughout Periods 1–4. Post-ovulatory FPM average concentrations (Period 4) did not differ (P > 0.05) between the ALT-treated females and controls. FPM average concentration from both groups increased in Period 4 compared to Periods 1–3 (P ≤ 0.05). Females receiving the ALT treatment also had lower (P ≤ 0.05) FGM metabolite average concentrations than control females during ovarian suppression (suggesting adrenal suppression). Collective results suggest that ovarian response to gonadotropin treatment in the cheetah was improved following oral progestin administration due to the normative increase in estradiol following stimulation for these females compared with control. This treatment should lead to more effective timed assisted reproduction procedures for this species.

The vaginal microbiome as a predictor for outcome of in vitro fertilization with or without intracytoplasmic sperm injection: a prospective study.

The vaginal microbiome as a predictor for outcome of in vitro fertilization with or without intracytoplasmic sperm injection: a prospective study.

Tecnología y profesionalismo como herramientas que nos ayudan a mejorar los resultados en técnicas de reproducción asistida

En estos últimos años la demanda por los procedimientos de reproducción asistida se ha ido incrementando de manera considerable. Todo parece indicar que esta tendencia va a seguir por este mismo camino, lo que nos obliga a los profesionales del área reproductiva a estar actualizados en cuanto a los últimos avances en tecnología y preparación de personal, con la única finalidad de ofrecer a los pacientes mejores tasas de embarazo. El actual objetivo de la reproducción asistida es conseguir la gestación y que esta llegue a término tras la primera transferencia de un embrión único. Ello supone un gran reto, dado que implica el esfuerzo conjunto por conseguir que la mayoría de nuestras pacientes logren el ansiado bebe sano en casa. Por ello, desde hace unos años asumimos este compromiso y entendimos que la única manera de alcanzar este objetivo era utilizar todas las herramientas disponibles que la tecnología nos puede ofrecer en el ámbito reproductivo.

El uso de la tecnología Time-Lapse y algoritmo predictivo KIDScore 5 ayudan a mejorar la selección de embriones euploides para la transferencia

Objetivos. Evaluar si el uso del algoritmo KIDScore 5 (known implantation data) puede ayudar a seleccionar entre los embriones euploides, para mejorar las tasas de embarazo e implantación en pacientes sometidas a procedimientos de reproducción asistida. Métodos. Estudio de cohorte retrospectivo en una clínica de fertilidad, desde octubre 2016 a diciembre 2018. Se estudió 1 049 embriones provenientes de 328 pacientes. Todos los embriones fueron cultivados en la incubadora Time-Lapse, Embryoscope® (Vitrolife®, Canadá) durante 5 a 6 días. De estos, 896 embriones (85,4%) fueron biopsiados y analizados mediante NGS y recibieron una valoración otorgada por el algoritmo predictivo KIDScore 5 (Vitrolife®, Canadá). Los 153 embriones restantes (14,6%) únicamente recibieron la valoración mediante el algoritmo predictivo KIDScore 5. Se realizó 256 transferencias únicas de embriones euploides en parejas sometidas a tratamientos de FIV en el laboratorio de reproducción asistida de la Clínica Inmater, Lima – Perú. Resultados. La tasa de implantación de los embriones euploides transferidos con valores de KIDScore = 6 versus los transferidos con valores de KIDScore = 1 tuvo diferencia estadísticamente signficativa (73,5% vs. 50,8%; p=0,030). Al evaluar la relación entre la tasa de euploidia embrionaria versus el resultado del valor de KIDScore 5, se obtuvo diferencias altamente significativas en las tasas de euploidia en los embriones con resultados de KIDScore 6 y 5 versus los de KIDScore 0 y 1 (60,5% vs. 45,7%; p=0,0004). Conclusiones. La selección embrionaria con ayuda del algoritmo KIDScore 5 ofrece ventaja en las tasas de implantación y embarazo únicamente cuando se transfieren embriones euploides. Su uso como criterio adicional a la selección embrionaria debiera ser considerado siempre que se acompañe estudio genético a los embriones a transferir. Los embriones euploides con valor más alto en la escala del algoritmo KIDScore 5, tienen mejores tasas de implantación y euploidía que los embriones con el valor mínimo de dicho algoritmo.

Study of clinical experience with different approaches to controlled ovarian hyperstimulation: a focus on safety and efficacy

ObjectivesCurrent retrospective cohort study analyses clinical database records of 4792 assisted reproduction procedures to assess the significance of target effectiveness endpoints from a safety perspective.MethodsStimulation protocols with urinary, recombinant or...

Effect of using frozen-thawed bovine semen contaminated with lumpy skin disease virus on in vitro embryo production.

Lumpy skin disease (LSD) is an important transboundary animal disease of cattle with significant economic impact because of the implications for international trade in live animals and animal products. LSD is caused by a Capripoxvirus, LSD virus (LSDV), and results in extensive hide and udder damage, fever and pneumonia. LSDV can be shed in semen of infected bulls for prolonged periods and transmitted venereally to cows at high doses. This study examined the effects of LSDV in frozen-thawed semen on in vitro embryo production parameters, including viral status of media and resulting embryos. Bovine oocytes were harvested from abattoir-collected ovaries and split into three experimental groups. After maturation, the oocytes were fertilized in vitro with frozen-thawed semen spiked with a high (HD) or a lower (LD) dose of LSDV, or with LSDV-free semen (control). Following day 7 and day 8 blastocyst evaluation, PCR and virus isolation were performed on all embryonic structures. After completing sufficient replicates to reach 1,000 inseminated oocytes, further in vitro fertilization (IVF) runs were performed to provide material for electron microscopy (EM) and embryo washing procedures. Overall, in vitro embryo yield was significantly reduced by the presence of LSDV in frozen-thawed semen, irrespective of viral dose. When semen with a lower viral dose was used, significantly lower oocyte cleavage rates were observed. LSDV could be detected in fertilization media and all embryo structures, when higher doses of LSDV were present in the frozen-thawed semen used for IVF. Electron microscopy demonstrated LSDV virions inside blastocysts. Following the International Embryo Transfer Society washing procedure resulted in embryos free of viral DNA; however, this may be attributable to a sampling dilution effect and should be interpreted with caution. Further research is required to better quantify the risk of LSDV transmission via assisted reproductive procedures.

Double-stranded sperm DNA damage is a cause of delay in embryo development and can impair implantation rates

To analyze the effect of single- and double-stranded sperm DNA fragmentation (ssSDF and dsSDF) on human embryo kinetics monitored under a time-lapse system. Observational, double blind, prospective cohort study. University spin-off and private center. One hundred ninety-six embryos from 43 infertile couples were included prospectively. None. SsSDF and dsSDF were analyzed in the same semen sample used for intracytoplasmic sperm injection. Embryo kinetics was then monitored using time-lapse technology, and the timing of each embryo division was obtained. When comparing embryos obtained from semen samples with low dsSDF and high dsSDF, splitting data using a statistically significant delay in high dsSDF was observed in second polar body extrusion, T4, T8, morula, and starting blastocyst and embryo implantation rates were impaired. Embryo kinetics and implantation rates are not significantly affected when high values of ssSDF are present. Different patterns of delay in embryo kinetics were observed for these different types of DNA damage: dsSDF caused a delay along all stages of embryo development; however, its major effect was observed at the second polar body extrusion and morula stages, coinciding with embryo DNA damage checkpoint activation as described before; ssSDF had its major effect at the pronucleus stage, but embryo kinetics was then restored at all following stages. The results show that dsSDF could be the main type of DNA damage that affects embryo development in intracytoplasmic sperm injection cycles, probably due to motility-based sperm selection in this assisted reproduction procedure. Double-stranded sperm DNA damage caused a delay in embryo development and impaired implantation, while single-stranded DNA damage did not significantly affect embryo kinetics and implantation.

Assisted reproductive technology: prevalence and associated factors in Southern Brazil.

ABSTRACTOBJECTIVE:To assess the prevalence of successful assisted reproductive technology and to identify the associated factors.METHODS:This population-based birth cohort study was carried out with 4,333 pregnant women expected to deliver in 2015 in the urban area of Pelotas, Southern Brazil. Use of an assisted reproductive technology procedure, type of assisted reproductive technology [in vitro fertilization or intracytoplasmic sperm injection or artificial insemination], number of embryos transferred, success of embryo transfer, number of attempts, and reported reasons for seeking assisted reproductive technology were the main outcomes measured. Use of an assisted reproductive technology procedure was analyzed according to sociodemographic, nutritional, reproductive history, and behavioral characteristics. Unadjusted and adjusted analyses were performed by logistic regression.RESULTS:Among the 4,275 newborns enrolled in the Pelotas 2015 Birth Cohort Study, 18 births (0.4%) were conceived by assisted reproductive technology. Most cases of assisted reproductive technology were by in vitro fertilization (70.6%). All cycles were performed in private clinics under direct out-of-pocket payment. Even after controlling for confounders, maternal age > 35 years, nulliparity and high family monthly income were strongly associated with assisted reproductive technology.CONCLUSIONS:The use of assisted reproductive technology services was reported by only a few women in the Pelotas 2015 Birth Cohort Study. Our study highlights sociodemographic factors associated to assisted reproductive technology procedures. To better understand the patterns and barriers in overall use of assisted reproductive technology services over time, national-level trend studies in assisted reproductive technology treatments and outcomes, as well as studies exploring the characteristics of women who have sought this kind of treatment are needed in low-middle income countries.

Code of Ethics and Conduct of the Brazilian Association of Embryologists - PRONÚCLEO

For more than three decades, Brazilian Clinical Embryologists have been working without specific regulations and following the standards adopted by other healthcare professionals. This document aims to guide behavior and decision-making, while providing directions to embryologist with the purpose of aiding professionals involved with assisted reproduction procedures and their patients. The Code of Ethics and Conduct is an important breakthrough and the first step toward regulating Clinical Embryology as a profession.

DNA methylation, environmental exposures and early embryo development.

The first crucial step in the developmental program occurs during pre-implantation, the time after the oocyte has been fertilized and before the embryo implants in the uterus. This period represents a vulnerable window as the epigenome undergoes dynamic changes in DNA methylation profiles. Alterations in the early embryonic reprogramming wave can impair DNA methylation patterns and induce permanent changes to the developmental program, leading to the onset of adverse health outcomes in offspring. Although there is an increasing body of evidence indicating that harmful exposures during pre-implantation embryo development can trigger lasting epigenetic alterations in offspring, the mechanisms are still not fully understood. Since physiological or pathological changes in DNA methylation can occur as a response to environmental cues, proper environmental milieu plays a critical role in the success of embryonic development. In this review, we depict the mechanisms behind the embryonic epigenetic reprogramming of DNA methylation and highlight how maternal environmental stressors (e.g., alcohol, heat stress, nutrient availability) during pre-implantation and assisted reproductive technology procedures affect development and DNA methylation marks.

Re: An overview of assisted reproductive technology procedures

Re: An overview of assisted reproductive technology procedures

Genes for anti-Müllerian hormone and androgen receptor are underexpressed in human cumulus cells surrounding morphologically highly graded oocytes.

Objectives:The aim of this study was to investigate the expression of genes crucial for the quality of the oocyte and whether expression levels of these genes in cumulus cells can be biological markers for the quality of the oocyte, zygote or embryo, or even for achievement of pregnancy after the assisted reproductive technology procedure. We examined the expression profile of the anti-Müllerian hormone (AMH) gene and its respective receptors: anti-Müllerian hormone receptor type 2 (AMHR2), follicle-stimulating hormone receptor (FSHR) and androgen receptor (AR) in cumulus cells (CCs) surrounding the oocyte, as well as AMH concentrations in follicular fluid of the associated follicle. The obtained gene expression levels were correlated with the morphological quality of the associated oocyte, zygote and embryo as well as with assisted reproductive technology outcome following the intracytoplasmic sperm injection procedure.Methods:This study involved 129 cumulus cells and 35 follicular fluid samples, taken from 58 patients undergoing the intracytoplasmic sperm injection procedure. Oocytes, zygotes and embryos were assessed for morphological quality. The relative gene expression of AMH, AMHR2, FSHR and AR was calculated using the delta–delta Ct method. Anti-Müllerian hormone concentrations in follicular fluids were measured by enzyme-linked immunosorbent assay.Results:The results yielded suggest a relationship between AMH, AR and oocyte morphology: AMH and AR gene expression levels in CCs surrounding morphologically optimal oocytes were significantly lower than in CCs surrounding oocytes with suboptimal morphology (p = 0.011 and p = 0.008, respectively). Statistically significant positive correlation was found between mRNA expression levels of AMH and FSHR (p < 0.001), AMH and AR (p = 0.001), AMHR2 and FSHR (p < 0.001), AMHR2 and AR (p < 0.001), as well as between FSHR and AR (p < 0.001).Conclusion:Assessed results point to AMH and AR relation with oocyte maturity, but not with its fertilization potential, or with embryo quality.

Pravo na slobodu kretanja zbog lečenja - primer "reproduktivnog turizma" u EU

Caring for the coming generations is vital for every society, and the processes that accompany human reproduction are of particular interest to parents, prenatal care providers and the public in general. The ethical and legal issues that arise in terms of new biological findings and their implementation in the field of assisted reproduction are set as a priority requirement in order to identify the values and interests that the legal order seeks to protect, prevent potential abuses, and clearly regulate the doctor-patient relationship in the context of medically assisted reproduction procedures. Although these days every EU country (except Ireland) has legislation governing assisted reproduction, accompanied by professional guidelines, there is still no European consensus on this matter. Moreover, it seems that the EU has so far refrained from adopting acts that will be binding on all Member States, consciously leaving this area largely dependent on the moral and ethical views of the Member States, which need to regulate it themselves. Due to the diversity of legal solutions in this area, as well as the ability to move freely through EU countries, a large number of patients decide to travel to countries that have more liberal legal solutions in the domain of assisted reproduction. This widespread phenomenon on the EU soil is called reproductive tourism and it represents a direct consequence of the lack of supranational legal sources in this area. It is through the example of reproductive tourism that the author analyses the extent to which the right to freedom of movement in the EU law affects the right of individuals, in Member States, to gain access to ethically controversial treatments available only in certain EU Member States.

Sperm mitochondrial DNA measures and semen parameters among men undergoing fertility treatment.

To examine associations between sperm mitochondrial DNA copy number (mtDNAcn), sperm mitochondrial DNA deletions (mtDNAdel), semen parameters and clinical infertility in an IVF setting. A total of 125 sperm samples were collected from men undergoing assisted reproductive procedures in an IVF clinic in Western Massachusetts, USA. Sperm mtDNAcn and mtDNAdel were measured by probe-based quantitative polymerase chain reaction. Semen parameters, clinical diagnoses of infertility, and infertility based on consecutive semen parameters, were fitted with mtDNAcn and mtDNAdel in linear models. The utility of sperm mtDNAcn and mtDNAdel to predict infertility was assessed by receiver operating characteristic curves. Adjusting for relevant covariates, both sperm mtDNAcn and mtDNAdel were associated with lower sperm concentration, count, motility and morphology (P ≤ 0.03). Sperm mtDNAcn and mtDNAdel were also associated with increased risks of clinical infertility based on current and consecutive semen samples. Sperm mtDNAcn had high predictive accuracy for consecutive diagnoses of clinical infertility (C-statistic: 0.91), whereas sperm mtDNAdel had moderate predictive accuracy (C-statistic: 0.75). Sperm mtDNAcn is a measure of consecutive abnormal semen parameters and has promise as a diagnostic test.

Freezing, Vitrification, and Freeze-Drying of Equine Spermatozoa: Impact on Mitochondrial Membrane Potential, Lipid Peroxidation, and DNA Integrity

Maintaining the integrity of equine sperm subjected to preservation protocols is essential for the successful development of assisted reproduction procedures. The aim of this study was to assess the mitochondrial membrane potential, lipid peroxidation, and DNA integrity of equine sperm subjected to freezing, vitrification, and freeze-drying. Eight ejaculates obtained from four Colombian Creole horses were subjected to programmable freezing, vitrification, and freeze-drying. After thawing or rehydration, sperm motility and kinetics were assessed through a CASA system. The mitochondrial membrane potential (ΔΨM), lipid peroxidation (LPO), and DNA fragmentation index (DFI) of the spermatozoa were assessed by flow cytometry using the DiOC6 (3), C11-Bodipy 581/591, and propidium iodide (PI) fluorescent dyes. The statistical analysis was conducted via generalized linear models, mean comparisons via the Duncan test, and a principal component analysis. A higher rate of spermatozoa with a high ΔΨM was found for freeze-drying (40.26 ± 7.79%) compared with freezing (21.82 ± 5.38%) and vitrification (5.32 ± 1.17%) (P < .05). Likewise, a higher rate of nonperoxidized viable spermatozoa (Bodipy-/PI-) was found for freeze-drying (35.98 ± 7.01%) in relation to frozen (10.34 ± 2.69%) and vitrified (7.07 ± 2.00%) sperm (P < .05). The DFI of vitrified spermatozoa (0.12 ± 0.04%) was higher when compared with the frozen (0.03 ± 0.01%) and freeze-dried (0.02 ± 0.01%) samples (P < .05). The researchers conclude that vitrification generates greater sperm alterations than freeze-drying and freezing, whereas freeze-drying produces lower LPO and higher ΔΨM for equine spermatozoa.

Numerical simulation of embryo transfer: how the viscosity of transferred medium affects the transport of embryos

BackgroundEmbryo transfer (ET) is a key step of assisted reproductive procedures, where the transferred medium containing the embryos is injected into the uterine cavity through a transcervical catheter and blended with intrauterine fluid in the uterine cavity. This procedure determines the delivery sites of embryos in the uterine cavity and has crucial impact on the implantation. Due to practical restrictions and ethical issues, it is often difficult to perform an in vivo study in humans to examine factors that affect the motions and delivery of embryos during ET. Alternatively, mathematical modeling is a powerful tool to that end.ResultsA computational model is developed to simulate the intrauterine mixing flow and track the embryo motions. Two important factors affecting the intrauterine flow are studied via this model: the viscosity of the transferred medium and the injection speed. Numerical results show that the dispersion pattern and the final delivery sites of the embryos are significantly influenced by the viscosity of the transferred medium. Specially, increasing the transferred medium viscosity close to that of the uterine fluid can enhance the probability that the embryos are delivered close to the fundus and keep them from being dragged backward to the cervix during catheter withdrawal. In addition, a slow injection speed can lower the driving force on the embryo during ET, which can prevent the embryo from being injured.ConclusionsBased on our study, the practice of using a transferred medium with similar viscosity to that of the uterine fluid and a slow injection speed is recommended for real embryo transfer procedures in clinic.

The prognostic role of thrombophilia in the treatment of infertility

Abstract Thrombophilia refers to a coagulation disorder that predisposes to thrombosis and thus increases the risk of thrombotic events. Both inherited and acquired thrombophilia are associated with vascular thrombosis and pregnancy-related complications, including infertility, recurrent miscarriage, and premature birth. Recently, thrombophilia has been increasingly encountered as an infertility factor, which gives the clinical relevance of the disease. The aim of the study was to investigate the prognostic role of thrombophilia in the treatment of infertility and the pregnancy of thrombophilic women during assisted reproduction procedures. Frequency of abortions increases and effectiveness of in vitro fertilization (IVF) decreases with age. Normal weight has a positive effect on assisted reproduction techniques (ART’s) outcome. Repeat IVF failure is more common in thrombophilia than in healthy women, and the “take home baby” ratio for IVF is 24%. Spontaneous abortion was most commonly observed in the PAI homozygous group followed by MTHFR homozygous mutation, MTHFR heterozygous mutation, and Factor V (Leiden) mutation. The most effective treatment was concomitant therapy with low molecular weight heparin and aspirin.

Knowledge gaps in male infertility: a reproductive endocrinology and infertility perspective.

Reproductive research has moved forward at a remarkable pace. Some of these advances are the result of a separation between male and female specialties, allowing focused study in specific areas of the field. However, the different training programs between male and female fertility specialists has created an environment in which some discoveries are not put in the greater context of clinical care. At times, interventions have been measured against surrogate markers of outcome that may not impact the most meaningful outcome for patients—the delivery of a healthy neonate. For example, medical and surgical interventions that use changes in semen parameters may have a limited impact on the likelihood of achieving a live birth due to the limitations inherent in the semen analysis for predicting outcomes. Other commonly used tests, such as sperm DNA fragmentation assays provide promising biological plausibility to account for subfertility of some male partners. However, until well defined thresholds for predicting outcomes in different treatment scenarios are available, changes in sperm DNA fragmentation testing is not an adequate outcome for measuring the utility of interventions. The biggest limitation for these tests remains their analysis of bulk semen. Tests allowing interrogation of the reproductive competence of a given sperm, while allowing that sperm to be used in assisted reproductive technology procedures remain elusive. Progress toward reaching this end (whether by hyaluronic acid binding, IMSI, or Ramen spectroscopy) is underway, but much remains to be learned. Achieving testing and capture of individual sperm would better facilitate studies that measure the most meaningful outcome for patients and providers—the delivery of a healthy baby.