Case records of the Massachusetts General Hospital. Case 11-2005. A 32-year-old pregnant woman with an abnormal fetal karyotype.
Explore the source record for details and available documents.
Adult↗
Biomedical subjects
Catherine Racowsky
Publications and source records attributed to Catherine Racowsky.
In vitro maturation of human oocytes and cumulus cells using a co-culture three-dimensional collagen gel system.
BACKGROUND: Deficiencies remain in the ability of in vitro-matured human oocytes to acquire full developmental competence and give rise to a healthy pregnancy. A clear deficiency of current systems utilizing human oocytes has been the absence of cumulus cells. In the present study, a three-dimensional (3D) co-culture system exploiting an extracellular matrix was developed and compared to conventional methods for its ability to support maturation of human oocytes. METHODS AND RESULTS: Cumulus cells were embedded into a 3D collagen gel matrix with individual oocytes added to each gel. Oocytes from the same patient cultured in the gel matrix matured to metaphase II at rates similar to those of cumulus-free oocytes cultured in individual microdrops. Following maturation of oocytes and fixation of intact gels, chromatin and cytoskeletal elements were assessed in oocytes and cumulus cells. The activities of the key cell cycle kinases, maturation-promoting factor (MPF) and mitogen-activated protein kinase (MAPK), were compared in oocytes matured under the two culture conditions. Compared with denuded oocytes, co-cultured oocytes exhibited increased MAPK activity, but no difference in MPF levels. CONCLUSIONS: This work characterizes a novel and efficacious culture system that takes advantage of the unique properties of the extracellular matrix, a 3D microenvironment, and the presence of cumulus cells for maturing human oocytes in vitro.
Adult↗
Aneuploidy involving chromosome 1 in failed-fertilized human oocytes is unrelated to maternal age.
PURPOSE: To study whether maternal meiotic errors in failed-fertilized oocytes involving chromosome 1 occur at frequencies similar to those involving other autosomes, and whether their frequency is affected by maternal age. METHODS: Using fluorescence in situ hybridization (FISH), frequencies of aneusomy and chromatid pre-division involving chromosomes 1, 16, 18, and 21 were determined for 273 failed-fertilized oocytes. RESULTS: The aneuploidy rate for chromosome 1 was 15.8%, and was neither age-dependent nor significantly different from that for chromosomes 16, 18 or 21. Only chromosome 16 exhibited an age-dependent increase in aneusomy rates. The frequency of chromatid pre-division was lower for chromosome 1 than for chromosome 18 (11.9% vs. 25.4%; p = 0.01), but not different from that for chromosomes 16 or 21. CONCLUSION: Aneuploidy involving chromosome 1 in failed-fertilized oocytes is unrelated to maternal age and occurs at a frequency similar to that for chromosomes 16, 18, and 21.
Aneuploidy↗
Optimum number of embryos to transfer in women more than 40 years of age undergoing treatment with assisted reproductive technologies.
OBJECTIVE: To determine whether increasing the number of embryos transferred beyond five increases pregnancy rates in women aged > 40 years. DESIGN: Retrospective analysis of cycles performed between January 1998 and July 2003. SETTING: University-affiliated teaching hospital. PATIENT(S): Women aged > 40 years undergoing a fresh cycle with a day-3 ET (n = 863). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Pregnancy, chemical pregnancy, miscarriage rates, number of viable fetuses at 12 weeks' gestation, live birth rates, and number of babies delivered. RESULT(S): Compared with patients with fewer than five embryos transferred, those having five or more embryos transferred had significantly increased pregnancy rates and live birth rates, more viable fetuses at 12 weeks, and significantly decreased miscarriage rates. None of these outcome variables differed between the five-embryo and more-than-five-embryo groups. There were no differences in outcome when only five embryos were transferred, regardless of whether five or more than five embryos were available. The number of embryos transferred did not significantly influence multiple birth rates. CONCLUSION(S): The present study demonstrates that in women aged > 40 years, five embryos is the optimum number to transfer, and transferring more than five does not confer any additional benefit to clinical outcome.
Adult↗
Embryo quality based on ovulation induction: defining the differences.
Patients undergoing IVF, with or without intracytoplasmic sperm injection, were treated with either recombinant human FSH or urine-derived FSH. Response to ovarian stimulation was monitored by ultrasound examinations and measurement of serum oestradiol concentrations. To define any differences in embryo quality and hence assisted reproductive technology success rates, a retrospective analysis of 811 recombinant FSH versus 555 urinary FSH cycles was undertaken. Embryo quality was assessed as embryo cell number and degree of fragmentation. Implantation and ongoing pregnancy rates were also compared. Use of recombinant FSH resulted in a higher percentage of mature oocytes, improved embryo cleavage, with more embryos available for freezing and higher implantation rates compared with urinary FSH. Oocyte and embryo quality were superior when recombinant FSH was used for ovarian stimulation compared with urinary FSH.
Adult↗
Assessment and optimization of oocyte quality during assisted reproductive technology treatment.
The health of fetuses, neonates, and adults depends on normal development of the oocytes and embryos from which they arise. In addition, the ability of an embryo to implant, sustain a pregnancy, and give rise to a viable offspring is unquestionably rooted in oogenesis. Therefore, defining markers that can reliably predict the best quality oocytes will prove invaluable in the care and management of infertility patients. Furthermore, identification of the best quality oocyte will also permit the transfer of a single embryo, thereby increasing overall pregnancy rates and reducing multiple rates. Although a few potentially important predictors of oocyte quality have been identified, their application for routine use in the assisted reproductive technology (ART) laboratory remains to be established. With the critical need for markers to be noninvasive, reliable, and assayable with a rapid turnaround, key areas that will take advantage of recent technological advances are identified in this article. Oocyte quality also needs to be improved in the management of human infertility by ART programs. Given that mechanisms determining oocyte quality are multifactorial and complex, several possible areas that continue to necessitate optimization are discussed.
Animals↗
Fluorescence in situ hybridization and spectral imaging analysis of human oocytes and first polar bodies.
We investigated the frequencies of abnormalities involving either chromosome 1, 16, 18, or 21 in failed-fertilized human oocytes. Although abnormalities involving chromosome 16 showed an age-dependent increase, results for the other chromosomes did not show statistically significant differences among the three age groups, <35 years, 35-39 years, and >39 years. The scoring of four chromosomes is likely to underestimate the true rate of aneuploid cells. Therefore, for a pilot study investigating a more-comprehensive analysis of oocytes and their corresponding first polar bodies, we developed a novel eight-probe chromosome enumeration scheme using fluorescence in situ hybridization and spectral imaging analysis.
Adult↗
The role of preparatory cycles in ovum donation recipients: a retrospective study.
PURPOSE: To determine whether preparatory cycles affect in vitro fertilization (IVF) outcome in ovum donation. METHODS: Medical records of 98 ovum donation recipients undergoing their first egg donation cycle were analyzed retrospectively. Preparatory cycles were performed in 50 patients with leuprolide acetate, estrogen, and progesterone. An endometrial biopsy was performed on day 10-12 of progesterone supplementation, to determine adequacy of the preparatory cycle. RESULTS: Pregnancy rates in women with and without preparatory cycles were 42.0% and 43.8%, respectively. Among ovum donation recipients who underwent preparatory cycles, the percentages of adequate endometrial biopsies in pregnant versus nonpregnant groups were 76.2% and 84.3%, respectively. No results showed statistical significance. CONCLUSIONS: Preparatory cycles do not increase pregnancy rates in ovum donation recipients. In addition, no correlation was noted between adequate endometrial biopsies and higher pregnancy rates.
Adult↗
Predictive usefulness of cycle day 10 follicle-stimulating hormone level in a clomiphene citrate challenge test for in vitro fertilization outcome in women younger than 40 years of age.
OBJECTIVE: To determine the usefulness of the day 10 FSH level of a clomiphene citrate challenge test in predicting IVF outcome in women younger than 40 years of age. DESIGN: Retrospective chart review. SETTING: Academic fertility center. PATIENT(S): Three hundred fifty-three women younger than 40 years of age undergoing 483 IVF cycles. MAIN OUTCOME MEASURE(S): Delivery rates. RESULT(S): An inverse relationship was observed between the likelihood of successful pregnancy and both cycle day 3 and cycle day 10 FSH levels. In women with a normal day 3 FSH level, elevated day 10 FSH levels predicted similarly poor IVF outcomes as for women with an abnormal day 3 FSH level. There was no specific threshold FSH level beyond which pregnancies failed to occur. Successful pregnancies were achieved with aggressive stimulation even in women with FSH levels greater than 20 mIU/mL. CONCLUSION(S): The clomiphene citrate challenge test (CCCT) correlates with IVF outcomes in women younger than 40 years of age. Stimulated day 10 FSH levels are strongly predictive of decreased IVF success even when day 3 FSH levels are normal. Results of the CCCT are useful for patient counseling before the IVF cycle and for choosing the optimal gonadotropin regimen.
Adult↗
Distinct microtubule and chromatin characteristics of human oocytes after failed in-vivo and in-vitro meiotic maturation.
BACKGROUND: While a complete failure of meiotic maturation following hCG administration is rare during IVF cycles, cases arise in which patients repeatedly display a high incidence of failure to complete maturation to metaphase II (MII) in vivo. For the immature oocytes of such patients, our objectives were (i) to ask whether progression to MII could be supported in vitro, and (ii) to define their microtubule/chromatin properties following in-vitro maturation (IVM). Together, these studies were aimed at augmenting our understanding of factors underlying meiotic arrest in the human. METHODS: Cases are presented here for two patients (A and B) producing oocytes that recurrently showed the inability to mature to metaphase II in vivo. Following IVM attempts, chromatin and microtubule characteristics were identified in those oocytes that remained arrested during meiosis I. RESULTS: In patient A, meiotically arrested oocytes exhibited clear defects in spindle and chromatin arrangements. In contrast, the majority of oocytes from patient B displayed normal MI and MII spindles with aligned chromosomes, although some oocytes exhibited indications for possible defects in cell cycle control. CONCLUSIONS: Together, these analyses illustrate two cases with oocytes exhibiting a common gross defect, that is meiotic maturation arrest, but revealing different aetiologies or manifestations as evidenced by the presence or absence of abnormal spindle/chromatin organization. This work reinforces the existence of intrinsic defects in oocytes of some patients, the molecular and cellular bases of which merit further investigation.
Cells, Cultured↗
The development of fertilized human ova to the blastocyst stage in KSOM(AA) medium: is a two-step protocol necessary?
Current protocols for the culture of human zygotes to blastocysts use two-step sequential media systems. The efficacy of a one-step system involving potassium simplex optimized medium (KSOM(AA)) has been investigated. In study 1, development of zygotes from days 1 to 3 in KSOM(AA) was compared with that for medium P-1. In study 2, embryos were cultured from days 1 to 3 in P-1 followed by culture from days 3 to 5 either in KSOM(AA) or medium CCM. In study 3, the ability of KSOM(AA) to support development of embryos from days 1 to 5, without medium renewal, was compared with the sequential media system P-1-->CCM. The cell numbers and fragmentation scores of day 3 embryos were distributed similarly following culture in KSOM(AA) or P-1. Significantly more KSOM(AA) embryos exhibited cytoplasmic pitting. Blastocyst formation rates were not significantly different whether embryos were cultured in the P-1-->KSOM(AA) or the P-1-->CCM systems, or when cultured from days 1 to 5 in KSOM(AA) without medium renewal or in P-1-->CCM. Five babies have been born from nine blastocysts transferred after extended culture in KSOM(AA). A one-step protocol involving KSOM(AA) can be used successfully to culture human zygotes to the blastocyst stage.
Blastocyst↗
Day 3 and day 5 morphological predictors of embryo viability.
Controlling multiple pregnancies in patients undergoing artificial reproductive procedures requires consideration of single embryo transfers. Therefore, refinements for embryo evaluation are needed that select for the most developmentally competent embryo. The present study was designed to identify day 3 and day 5 morphological predictors of viability following transfers in which the morphology and fate of each embryo was precisely determined. Assessments on day 3 included cell number, and the extent of fragmentation and asymmetry, and on day 5, the developmental stage. Embryos resulting in a viable fetus at 11 weeks gestation were considered developmentally competent. The relationships among individual and collective embryo morphological characteristics were evaluated. Analysis of the interactions among morphological characteristics of embryos transferred on day 3 enabled identification of a multivariable selection order. Assessment of day 5 embryos revealed that expanding and expanded blastocysts exhibited comparable developmental potential that was superior to that of either morulae or early blastocysts. However, expanding or expanded blastocysts derived from 7-cell or 8-cell embryos were developmentally superior to those derived from other cleavage stages, regardless of fragmentation or asymmetry. Collectively, these findings further understanding of morphological predictors of viability, thereby improving the ability to select the most viable embryo for transfer.
Adult↗