Growth and differentiation factor 9 promotes oocyte growth at the primary but not the early secondary stage in three-dimensional follicle culture.

PurposeFactors that differentially regulate oocyte and granulosa cell growth within the early preantral follicle and how these factors differ at each stage of follicle growth remain poorly understood. The aim of this study was to isolate and evaluate the effect of recombinant growth and differentiation factor 9 (GDF9) on oocyte and granulosa cell growth at the primary and early secondary stages of preantral follicle growth during in vitro culture.MethodsPrimary stage follicles (diameters of 50-89 μm) and early secondary stage follicles (diameters of 90-120 μm) were isolated from immature mice, and individual, intact follicles were cultured in vitro in the presence and absence of recombinant GDF9. The effects of GDF9 on follicle growth were determined by the assessment of changes in the follicle volume during culture. The growth of the granulosa cell and oocyte compartments of the follicles was evaluated separately at each stage.ResultsGDF9 significantly increased the growth of isolated follicles at both the primary and early secondary follicle stages. Independent evaluation of the granulosa cell and oocyte compartments revealed that, while GDF9 promoted granulosa cell growth at both stages of folliculogenesis, oocyte growth was stage specific. GDF9 promoted growth of the oocyte at the primary, but not the early secondary, follicle stage.ConclusionsThese findings demonstrate a stage-specific role for GDF9 in the regulation of oocyte and granulosa cell growth at the primary and early secondary stages of preantral follicle development

Growth and differentiation factor 9 promotes oocyte growth at the primary but not the early secondary stage in three-dimensional follicle culture.

PurposeFactors that differentially regulate oocyte and granulosa cell growth within the early preantral follicle and how these factors differ at each stage of follicle growth remain poorly understood. The aim of this study was to isolate and evaluate the effect of recombinant growth and differentiation factor 9 (GDF9) on oocyte and granulosa cell growth at the primary and early secondary stages of preantral follicle growth during in vitro culture.MethodsPrimary stage follicles (diameters of 50-89 μm) and early secondary stage follicles (diameters of 90-120 μm) were isolated from immature mice, and individual, intact follicles were cultured in vitro in the presence and absence of recombinant GDF9. The effects of GDF9 on follicle growth were determined by the assessment of changes in the follicle volume during culture. The growth of the granulosa cell and oocyte compartments of the follicles was evaluated separately at each stage.ResultsGDF9 significantly increased the growth of isolated follicles at both the primary and early secondary follicle stages. Independent evaluation of the granulosa cell and oocyte compartments revealed that, while GDF9 promoted granulosa cell growth at both stages of folliculogenesis, oocyte growth was stage specific. GDF9 promoted growth of the oocyte at the primary, but not the early secondary, follicle stage.ConclusionsThese findings demonstrate a stage-specific role for GDF9 in the regulation of oocyte and granulosa cell growth at the primary and early secondary stages of preantral follicle development

An Economic Analysis of Cell-Free DNA Non-Invasive Prenatal Testing in the US General Pregnancy Population

<div><p>Objective</p><p>Analyze the economic value of replacing conventional fetal aneuploidy screening approaches with non-invasive prenatal testing (NIPT) in the general pregnancy population.</p><p>Methods</p><p>Using decision-analysis modeling, we compared conventional screening to NIPT with cell-free DNA (cfDNA) analysis in the annual US pregnancy population. Sensitivity and specificity for fetal aneuploidies, trisomy 21, trisomy 18, trisomy 13, and monosomy X, were estimated using published data and modeling of both first- and second trimester screening. Costs were assigned for each prenatal test component and for an affected birth. The overall cost to the healthcare system considered screening costs, the number of aneuploid cases detected, invasive procedures performed, procedure-related euploid losses, and affected pregnancies averted. Sensitivity analyses evaluated the effect of variation in parameters. Costs were reported in 2014 US Dollars.</p><p>Results</p><p>Replacing conventional screening with NIPT would reduce healthcare costs if it can be provided for $744 or less in the general pregnancy population. The most influential variables were timing of screening entry, screening costs, and pregnancy termination rates. Of the 13,176 affected pregnancies undergoing screening, NIPT detected 96.5% (12,717/13,176) of cases, compared with 85.9% (11,314/13,176) by conventional approaches. NIPT reduced invasive procedures by 60.0%, with NIPT and conventional methods resulting in 24,596 and 61,430 invasive procedures, respectively. The number of procedure-related euploid fetal losses was reduced by 73.5% (194/264) in the general screening population.</p><p>Conclusion</p><p>Based on our analysis, universal application of NIPT would increase fetal aneuploidy detection rates and can be economically justified. Offering this testing to all pregnant women is associated with substantial prenatal healthcare benefits.</p></div

Aneuploidy incidence rates and performance of conventional screening approaches in the first and second trimester for a general population.

<p>^{^} Conventional screening based on maternal age, nuchal translucency, maternal serum pregnancy-associated plasma protein A [PAPPA] and free beta human chorionic gonadotropin [hCG] at 12 weeks gestational age.</p><p>* NIPT sensitivity and specificity was based on pooled data from 19 published studies (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0132313#pone.0132313.s003" target="_blank">S2 Table</a>); NIPT sensitivity and specificity was considered to be independent of pregnancy stage and maternal prior risk.</p><p>^† First trimester sensitivity for trisomy 13 screening is based on the algorithm developed for trisomy 18 screening [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0132313#pone.0132313.ref030" target="_blank">30</a>].</p><p>^‡ Second trimester sensitivity for trisomy 13 screening is assumed to be equal to the proportion of trisomy 13 affected pregnancies serendipitously identified as a false positive in Down syndrome and trisomy 18 screening.</p><p>^§ No specific screening protocols exist for trisomy 13 and monosomy X.</p><p>Aneuploidy incidence rates and performance of conventional screening approaches in the first and second trimester for a general population.</p

Model inputs and economic value of NIPT for fetal aneuploidy screening.

<p>FP(s), false positives; CVS, chorionic villus sampling; NIPT, non-invasive prenatal testing</p><p>^{^} NIPT values that correspond to the range applied to each input variable. These values can be compared to the $744 value assigned to the set of baseline model inputs.</p><p>^¥ Alternative scenario showing results when all initial screening is performed in the second trimester.</p><p>* Baseline invasive testing rates for true positives were 73$369) was a buildup of all the individual components (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0132313#sec007" target="_blank">Results</a>).</p><p>^§ The range of lifetime costs of an affected child was baseline ±20%; the upper and lower range values for each indication were modified together.</p><p>^˅Most of the variability is attributable to lifetime costs for Down syndrome (see text).</p><p>Model inputs and economic value of NIPT for fetal aneuploidy screening.</p

Incidence of X and Y Chromosomal Aneuploidy in a Large Child Bearing Population

<div><p>Background</p><p>X&Y chromosomal aneuploidies are among the most common human whole-chromosomal copy number changes, but the population-based incidence and prevalence in the child-bearing population is unclear.</p><p>Methods</p><p>This retrospective analysis of prospectively collected data leveraged a routine non-invasive prenatal test (NIPT) using parental genotyping to estimate the population-based incidence of X&Y chromosome variations in this population referred for NIPT (generally due to advanced maternal age).</p><p>Results</p><p>From 141,916 women and 29,336 men, 119 X&Y chromosomal abnormalities (prevalence: 1 in 1,439) were identified. Maternal findings include: 43 cases of 45,X (40 mosaic); 30 cases of 47,XXX (12 mosaic); 3 cases of 46,XX uniparental disomy; 2 cases of 46,XY/46,XX; 23 cases of mosaicism of unknown type; 2 cases of 47,XX,i(X)(q10). Paternal findings include: 2 cases of 47,XXY (1 mosaic); 10 cases of 47,XYY (1 mosaic); 4 partial Y deletions.</p><p>Conclusions</p><p>Single chromosome aneuploidy was present in one of every 1,439 individuals considered in this study, showing 47,XXX; 47,XX,i(X)(q10); 47,XYY; 47,XXY, partial Y deletions, and a high level of mosaicism for 45,X. This expands significantly our understanding of X&Y chromosomal variations and fertility issues, and is critical for families and adults affected by these disorders. This current and extensive information on fertility will be beneficial for genetic counseling on prenatal diagnoses as well as for newly diagnosed postnatal cases.</p></div

Maternal and Paternal Genotypes and Population Incidences.

<p>Maternal and Paternal Genotypes and Population Incidences.</p

Parental Aneuploidy & Corresponding Fetal NIPT Outcomes.

<p>Parental Aneuploidy & Corresponding Fetal NIPT Outcomes.</p