RESUMEN

OBJECTIVE: To compare autosomal and sex chromosome aneuploidy rates of embryos derived from sperm with abnormal and normal parameters. DESIGN: Retrospective cohort study. SETTING: Assisted reproduction center. PATIENT(S): Three thousand eight hundred thirty-five embryos generated from 629 couples undergoing IVF. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Incidence of aneuploidy in the trophectoderm of blastocyst embryos derived from standard IVF embryos and intracytoplasmic (ICSI) males with normal and oligozoospermic semen samples, in couples with donor eggs (mean maternal age, 25.0 years) and their own eggs (mean maternal age, 35.4 years). RESULT(S): The rate of sex chromosome aneuploidy was significantly (around threefold) higher in the oligozoospermic group compared with in both control groups (standard vs. ICSI insemination). This applied whether donor (young) or own (older) eggs were used. Significant differences were seen in the oligozoospermic samples for autosomes 1, 2, 11 (own eggs), and 18 (donor eggs) compared with both control groups; however, no significant difference was seen between each of the treatment groups for the overall rate of autosomal aneuploidy. No significant differences were seen between the two control groups (normozoospermic males, standard vs. ICSI insemination) in either of the egg group types for any chromosome pairs. CONCLUSION(S): Severe male factor infertility is associated with a significant increase in the occurrence of sex chromosome abnormalities in blastocyst embryos compared with in embryos derived from normal semen samples. Aneuploidy rates in embryos derived from sperm with normal parameters were not significantly different whether ICSI or standard insemination was used to achieve fertilization. These results highlight severe male factor infertility as a possible referral category for preimplantation comprehensive chromosomal screening.

Asunto(s)

Aneuploidia , Blastocisto , Análisis de Semen/efectos adversos , Aberraciones Cromosómicas Sexuales , Adulto , Blastocisto/metabolismo , Blastocisto/patología , Femenino , Fertilización In Vitro , Humanos , Infertilidad Masculina/genética , Masculino , Embarazo , Diagnóstico Preimplantación , Estudios Retrospectivos , Factores de Riesgo , Aberraciones Cromosómicas Sexuales/embriología , Aberraciones Cromosómicas Sexuales/estadística & datos numéricos , Espermatozoides/patología

RESUMEN

Crossover recombination reshuffles genes and prevents errors in segregation that lead to extra or missing chromosomes (aneuploidy) in human eggs, a major cause of pregnancy failure and congenital disorders. Here we generate genome-wide maps of crossovers and chromosome segregation patterns by recovering all three products of single female meioses. Genotyping >4 million informative SNPs from 23 complete meioses allowed us to map 2,032 maternal and 1,342 paternal crossovers and to infer the segregation patterns of 529 chromosome pairs. We uncover a new reverse chromosome segregation pattern in which both homologs separate their sister chromatids at meiosis I; detect selection for higher recombination rates in the female germ line by the elimination of aneuploid embryos; and report chromosomal drive against non-recombinant chromatids at meiosis II. Collectively, our findings show that recombination not only affects homolog segregation at meiosis I but also the fate of sister chromatids at meiosis II.

Asunto(s)

Segregación Cromosómica , Recombinación Genética , Células Cultivadas , Mapeo Cromosómico , Intercambio Genético , Femenino , Genoma Humano , Humanos , Meiosis , Oocitos/fisiología , Cuerpos Polares , Polimorfismo de Nucleótido Simple , Análisis de Secuencia de ADN

Asunto(s)

Citogenética , Haplotipos/genética , Síndrome de Marfan/diagnóstico , Diagnóstico Preimplantación , Biopsia , Femenino , Fertilización In Vitro , Humanos , Síndrome de Marfan/genética , Síndrome de Marfan/patología , Polimorfismo de Nucleótido Simple/genética , Embarazo

RESUMEN

Preimplantation genetic diagnosis (PGD) for monogenic disorders has the drawback of time and cost associated with tailoring a specific test for each couple, disorder, or both. The inability of any single assay to detect the monogenic disorder in question and simultaneously the chromosomal complement of the embryo also limits its application as separate tests may need to be carried out on the amplified material. The first clinical use of a novel approach ('karyomapping') was designed to circumvent this problem. In this example, karyomapping was used to confirm the results of an existing PGD case detecting both chromosomal abnormalities and a monogenic disorder (Smith-Lemli-Opitz [SLO] syndrome) simultaneously. The family underwent IVF, ICSI and PGD, and both polar body and cleavage stage biopsy were carried out. Following whole genome amplification, array comparative genomic hybridisation of the polar bodies and minisequencing and STR analysis of single blastomeres were used to diagnose maternal aneuploidies and SLO status, respectively. This was confirmed, by karyomapping. Unlike standard PGD, karyomapping required no a-priori test development. A singleton pregnancy and live birth, unaffected with SLO syndrome and with no chromosome abnormality, ensued. Karyomapping is potentially capable of detecting a wide spectrum of monogenic and chromosome disorders and, in this context, can be considered a comprehensive approach to PGD.

Asunto(s)

Trastornos de los Cromosomas/genética , Cariotipificación/métodos , Diagnóstico Preimplantación/métodos , Blastómeros/patología , Aberraciones Cromosómicas , Cromosomas/ultraestructura , Hibridación Genómica Comparativa/métodos , Análisis Mutacional de ADN , Femenino , Fertilización In Vitro , Humanos , Recién Nacido , Nacimiento Vivo , Masculino , Cuerpos Polares/patología , Embarazo , Resultado del Embarazo , Síndrome de Smith-Lemli-Opitz/diagnóstico , Síndrome de Smith-Lemli-Opitz/genética , Inyecciones de Esperma Intracitoplasmáticas/métodos

RESUMEN

Organisation of chromosome territories in interphase nuclei has been studied in many systems and positional alterations have been associated with disease phenotypes (e.g. laminopathies, cancer) in somatic cells. Altered nuclear organisation is also reported in developmental processes such as mammalian spermatogenesis where a "chromocentre" model is proposed with the centromeres and sex chromosomes repositioning to the nuclear centre. The purpose of this study was to test the hypothesis that alterations in nuclear organisation of human spermatozoa are associated with defects upstream in spermatogenesis (as manifest in certain infertility phenotypes). The nuclear address of (peri-) centromeric loci for 18 chromosomes (1-4, 6-12, 15-18, 20, X and Y) was assayed in 20 males using established algorithms for 3D extrapolations of 2D data. The control group comprised 10 fertile sperm donors while the test group was 10 patients with severely compromised semen parameters including high sperm aneuploidy. All loci examined in the control group adopted defined, interior positions thus providing supporting evidence for the presence of a chromocentre and interior sex chromosome territories. In the test group however there were subtle alterations in the nuclear address for certain centromeres in individual patients and, when all patient results were pooled, some different nuclear addresses were observed for chromosomes 3, 6, 12 and 18. Considering the extensive impairment of spermatogenesis in the test group (evidenced by compromised semen parameters and increased chromosome abnormalities), the observed differences in nuclear organisation for centromeric loci compared to the controls were modest. A defined pattern of nuclear reorganisation of centromeric loci in sperm heads therefore appears to be a remarkably robust process, even if spermatogenesis is severely compromised.

Asunto(s)

Núcleo Celular/genética , Centrómero/genética , Espermatozoides/ultraestructura , Adulto , Aberraciones Cromosómicas , Cromosomas Humanos Y , Fertilidad/genética , Humanos , Infertilidad Masculina/genética , Interfase/genética , Masculino , Persona de Mediana Edad , Espermatogénesis/genética

RESUMEN

Trisomy causes mental retardation, pregnancy loss, IVF failure, uniparental disomy and several other pathologies, and its accurate detection is thus clinically essential. Most trisomies arise at meiosis I and are associated with increasing maternal age and reduction or alteration in recombination patterns. Investigations into the relationship between trisomy and meiotic recombination have used short tandem repeat markers; however, this approach is limited by the resolution with which the position of crossovers can identified. As cytogenetics enters the post-genomic era, recent work has used array comparative genomic hybridisation (aCGH) to screen for trisomy of all 24 chromosomes, determining chromosome copy number by dosage analysis. However, aCGH has a fundamental drawback for studying the aetiology of trisomy since neither the parent and phase of origin nor uniparental disomy can be ascertained. The development of SNP microarrays has made it possible to analyse multiple loci for sequence variation, and the proprietary software provided can determine the presence of aneuploidy by algorithms based on fluorescence intensity. To the best of our knowledge, however, such software is not equipped to determine the phase of origin of the error or the position of any chiasmata. In this study, therefore, we present an algorithm to determine the parent of origin, the phase of origin and the location of chiasmata in a series of nine "trisomy triplets" (i.e. samples derived from father, mother and their trisomic foetus). Novel adaptations of well-established principles are applied along with a simple algorithm written in Microsoft Excel for visualisation of the results. Such analysis has a range of applications in preimplantation and prenatal diagnosis.

Asunto(s)

Algoritmos , Técnicas Genéticas , Polimorfismo de Nucleótido Simple , Intercambio de Cromátides Hermanas/genética , Trisomía/genética , Padre , Femenino , Genotipo , Humanos , Masculino , Madres , Embarazo , Complicaciones del Embarazo/diagnóstico , Complicaciones del Embarazo/genética

RESUMEN

The use of genome wide single nucleotide polymorphism (SNP) arrays for high resolution molecular cytogenetic analysis using a combination of quantitative and genotype analysis is well established. This study demonstrates that by Mendelian analysis of the SNP genotypes of the parents and a sibling or other appropriate family member to establish phase, it is possible to identify informative loci for each of the four parental haplotypes across each chromosome and map the inheritance of these haplotypes and the position of any crossovers in the proband. The resulting 'karyomap', unlike a karyotype, identifies the parental and grandparental origin of each chromosome and chromosome segment and is unique for every individual being defined by the independent segregation of parental chromosomes and the pattern of non-recombinant and recombinant chromosomes. Karyomapping, therefore, enables both genome wide linkage based analysis of inheritance and detection of chromosome imbalance where either both haplotypes from one parent are present (trisomy) or neither are present (monosomy/deletion). The study also demonstrates that karyomapping is possible at the single cell level following whole genome amplification and, without any prior patient or disease specific test development, provides a universal linkage based methodology for preimplantation genetic diagnosis readily available worldwide.

Asunto(s)

Mapeo Cromosómico/métodos , Enfermedades Genéticas Congénitas/diagnóstico , Enfermedades Genéticas Congénitas/genética , Estudio de Asociación del Genoma Completo/métodos , Haplotipos , Adulto , Niño , Cromosomas Humanos , Fibrosis Quística/genética , Regulador de Conductancia de Transmembrana de Fibrosis Quística/genética , Femenino , Genotipo , Humanos , Masculino , Padres , Linaje , Polimorfismo de Nucleótido Simple/genética

RESUMEN

It is now well known that levels of sperm disomy correlate to levels of infertility (as well as other factors). The risk of perpetuating aneuploidy to the offspring of infertile males undergoing intracytoplasmic sperm injection (ICSI) has become a hotly debated issue in assisted reproduction; however, there remain barriers to the practical implementation of offering sperm disomy screening in a clinical setting. The major barrier is the operator time taken to analyze a statistically meaningful (sufficient) number of cells. The introduction of automated 'spot counting' software-hardware combinations presents a potential solution to this problem. In this preliminary validation study, we analyzed 10 patients, both manually and using a commercially available spot counter. Results show a statistically significant correlation between both approaches for scoring of sperm disomy, but no correlation is found when scoring for diploid sperm. The most likely explanation for the latter is an apparent overscoring of two closely associated sperm heads as a single diploid cell. These results, and similar further studies that will ensue, help to inform cost-benefit analyses that individual clinics need to carry out in order to decide whether to adopt sperm aneuploidy screening as a routine tool for the assessment of sperm from men requiring ICSI treatment.

Asunto(s)

Aberraciones Cromosómicas , Espermatozoides/metabolismo , Automatización , Humanos , Hibridación Fluorescente in Situ , Masculino , Reproducibilidad de los Resultados

RESUMEN

In the field of nanotechnology, quantum dots (QDs) are a novel class of inorganic fluorochromes composed of nanometre-scale crystals made of a semiconductor material. Given the remarkable optical properties that they possess, they have been proposed as an ideal material for use in fluorescent in-situ hybridization (FISH). That is, they are resistant to photobleaching and they excite at a wide range of wavelengths but emit light in a very narrow band that can be controlled by particle size and thus have the potential for multiplexing experiments. The principal aim of this study was to compare the potential of QDs against traditional organic fluorochromes in both indirect (i.e. QD-conjugated streptavidin) and direct (i.e. synthesis of QD-labelled FISH probes) detection methods. In general, the indirect experiments met with a degree of success, with FISH applications demonstrated for chromosome painting, BAC mapping and use of oligonucleotide probes on human and avian chromosomes/nuclei. Many of the reported properties of QDs (e.g. brightness, 'blinking' and resistance to photobleaching) were observed. On the other hand, signals were more frequently observed where the chromatin was less condensed (e.g. around the periphery of the chromosome or in the interphase nucleus) and significant bleed-through to other filters was apparent (despite the reported narrow emission spectra). Most importantly, experimental success was intermittent (sometimes even in identical, parallel experiments) making attempts to improve reliability difficult. Experimentation with direct labelling showed evidence of the generation of QD-DNA constructs but no successful FISH experiments. We conclude that QDs are not, in their current form, suitable materials for FISH because of the lack of reproducibility of the experiments; we speculate why this might be the case and look forward to the possibility of nanotechnology forming the basis of future molecular cytogenetic applications.

Asunto(s)

Colorantes Fluorescentes/química , Hibridación Fluorescente in Situ/métodos , Nanotecnología/métodos , Puntos Cuánticos , Animales , Biotina/metabolismo , Biotinilación , Carbocianinas/metabolismo , Núcleo Celular/metabolismo , Células Cultivadas , Pollos , Pintura Cromosómica , Cromosomas/genética , Cromosomas Artificiales Bacterianos/genética , Cromosomas Humanos Par 12/genética , Células Clonales , ADN/metabolismo , Digoxigenina/metabolismo , Fluoresceína-5-Isotiocianato/metabolismo , Técnica del Anticuerpo Fluorescente Indirecta , Colorantes Fluorescentes/metabolismo , Humanos , Hibridación Genética , Indicadores y Reactivos/metabolismo , Indoles/metabolismo , Linfocitos/citología , Linfocitos/metabolismo , Masculino , Metafase , Microscopía Fluorescente , Sondas de Oligonucleótidos/química , Fotoblanqueo , Semiconductores , Espermatozoides/citología , Espermatozoides/metabolismo , Estreptavidina/metabolismo , Xantenos/metabolismo

Asunto(s)

Aneuploidia , Fertilización In Vitro , Pruebas Genéticas , Diagnóstico Preimplantación , Adulto , Trastornos de los Cromosomas/diagnóstico , Análisis Costo-Beneficio , Femenino , Pruebas Genéticas/métodos , Humanos , Edad Materna , Embarazo , Índice de Embarazo , Diagnóstico Preimplantación/economía

RESUMEN

Preimplantation genetic diagnosis for single gene disorders is usually performed using polymerase chain reaction (PCR)-based methodologies modified for use in single cells. At present, single cell PCR tests require costly and time-consuming development and validation of highly sensitive amplification strategies to cover a growing number of mutations responsible for genetic disease. Whole-genome amplification (WGA) provides an opportunity to amplify the genome from a single blastomere to a level at which multiple tests can be performed on the same cell. Early WGA methods (primer extension preamplification and degenerate oligonucleotide-primed PCR) have not proved sufficiently accurate and reliable for routine clinical use. However, WGA using multiple displacement amplification (MDA) offers approx 5 million-fold amplification with fidelity, apparently sidestepping the limitation of a single cell, and is sufficient for use in most off-the-shelf molecular tests. This chapter describes an optimized MDA protocol for the preparation of genomic DNA from single fibroblasts.

Asunto(s)

Genoma Humano/genética , Reacción en Cadena de la Polimerasa/métodos , Diagnóstico Preimplantación/métodos , Células/química , Regulador de Conductancia de Transmembrana de Fibrosis Quística/genética , Humanos , Repeticiones de Microsatélite

RESUMEN

Frozen-thawed embryos accounted for 39% (249 of 639) of live births from 931 consecutive first oocyte retrievals after median follow-up of 6.5 years with consistent use of pronuclear-stage freezing and cleavage-stage transfer. Survival after thaw was 95% (2,129 of 2,247). Implantation and live birth rates per individual frozen-thawed embryo transfered were 22% (431 of 1,937) and 18% (346 of 1,937), respectively.

Asunto(s)

Criopreservación/estadística & datos numéricos , Transferencia de Embrión/estadística & datos numéricos , Embrión de Mamíferos/patología , Infertilidad/epidemiología , Infertilidad/terapia , Nacimiento Vivo/epidemiología , Oocitos/patología , Adulto , Femenino , Estudios de Seguimiento , Calor , Humanos , Incidencia , Infertilidad/patología , Masculino , Minnesota/epidemiología , Oocitos/trasplante , Embarazo , Índice de Embarazo , Estudios Retrospectivos , Resultado del Tratamiento

RESUMEN

OBJECTIVE: To determine the effectiveness of a novel treatment designed to remove human immunodeficiency virus (HIV) -1 and hepatitis C virus (HCV) from spiked semen and to evaluate sperm motility and viability after treatment. DESIGN: A prospective clinical laboratory-based study. SETTING: The human studies were conducted in academic and national research environments. The bovine study was conducted in an accredited research facility. PATIENT(S): Healthy volunteers provided the semen samples used in the human studies; abattoir-derived material was used for the bovine embryo production study. INTERVENTIONS(S): None. MAIN OUTCOME MEASURE(S): Cytopathic, reverse transcriptase-polymerase chain reaction, and branched DNA assays were used to test the efficacy of the procedure for inactivating or removing viruses from spiked semen; standard semen evaluation criteria were used to assess the effects of the procedures on sperm motility and viability. RESULT(S): Trypsin exposure significantly reduced the infectivity of HIV-1. The triple density gradient treatment, with or without trypsin, had no detrimental affect on fresh or cryopreserved/thawed sperm 2-48 hours after treatment. The treatment of semen spiked with HIV-1 or HCV indicated that the procedure was effective for reducing viral copies to undetectable levels or below levels of clinical relevance. CONCLUSION(S): The procedure was effective for significantly inactivating or reducing HIV-1 and HCV in spiked semen without adversely affecting sperm quality.

Asunto(s)

VIH-1/efectos de los fármacos , Hepacivirus/efectos de los fármacos , Semen/efectos de los fármacos , Semen/virología , Motilidad Espermática/efectos de los fármacos , Tripsina/farmacología , Animales , Bovinos , Centrifugación por Gradiente de Densidad/métodos , Dinamarca , Femenino , VIH-1/fisiología , Hepacivirus/fisiología , Humanos , Masculino , Estudios Prospectivos , Quebec , Semen/fisiología , Motilidad Espermática/fisiología , Carga Viral/estadística & datos numéricos

RESUMEN

OBJECTIVE: We set out to characterize the expression of nine genes in human preimplantation embryos and determine whether abnormal morphology is associated with altered gene activity. DESIGN: Reverse transcription and real-time polymerase chain reaction were used to quantify the expression of multiple genes in each embryo. The genes studied have various important cellular roles (e.g., cell cycle regulation, DNA repair, and apoptosis). SETTING: Research laboratory working closely with a clinical IVF practice. PATIENT(S): Over 50 embryos were donated by infertile patients (various etiologies). Among these, all major stages of preimplantation development and a variety of common morphologic abnormalities were represented. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Quantification of mRNA transcripts. RESULT(S): We detected an association between certain forms of abnormal morphology and disturbances of gene activity. Cellular fragmentation was associated with altered expression of several genes, including TP53, suggesting that fragmenting blastomeres are suffering stress of a type monitored by p53, possibly as a consequence of suboptimal culture conditions. CONCLUSION(S): Appropriate gene expression is vital for the regulation of metabolic pathways and key developmental events. Our data indicates a possible causal relationship between changes in gene expression and the formation of clinically relevant abnormal embryo morphologies. We hypothesize that embryos with expression profiles characteristic of good morphology and appropriate for their developmental stage have the greatest potential for implantation. If confirmed, this could lead to a new generation of preimplantation genetic diagnosis (PGD) tests for assessing embryo viability and predicting implantation potential.

Asunto(s)

Blastocisto/fisiología , Embrión de Mamíferos/anomalías , Embrión de Mamíferos/metabolismo , Desarrollo Embrionario/genética , Regulación del Desarrollo de la Expresión Génica/fisiología , Análisis por Conglomerados , Humanos , Reacción en Cadena de la Polimerasa/métodos

RESUMEN

Glucose uptake was used as a measure of metabolic activity and implantation potential to compare vitrification and slow freezing in a prospective randomized trial using murine blastocysts. Frozen 2-cell embryos (n = 132) thawed and cultured for 48 h to the blastocyst stage were randomly divided into four groups: (i) control - not refrozen; (ii) slow freezing using a programmed rate (PR); (iii) vitrification by super-cooled (VSC) liquid nitrogen; and (iv) vitrification in liquid nitrogen (VLN). Upon re-thawing, embryos were cultured individually for 24 h to determine glucose uptake non-invasively. Morphological assessments included total cell counts and inner cell mass (ICM) detection following immunosurgery. Mean glucose uptake was lower for each treatment (PR and VSC, 4.3 pmol/embryo per h; VLN, 4.9 pmol/embryo per h) versus controls (6.8 pmol/embryo per h). PR and VSC embryos had fewer cells (57.4 +/- 24.2 and 64.1 +/- 31.5) versus controls (85.7 +/- 26.2), and fewer embryos containing a detectable ICM (42.9 and 61.8%) compared with controls (88.2%). The only difference between control and VLN embryos was absolute glucose uptake, although in both treatments glucose uptake was increased from embryos with an ICM compared with those without. Glucose uptake appears to be a sensitive, non-invasive method to validate cryopreservation protocols.

Asunto(s)

Blastocisto , Criopreservación/métodos , Glucosa/metabolismo , Animales , Blastocisto/metabolismo , Técnicas de Cultivo de Embriones , Desarrollo Embrionario , Femenino , Masculino , Ratones

RESUMEN

Insulin action is mediated by two insulin receptor (IR) isoforms, differing in mitogenic and metabolic function. IR isoform expression might occur in human granulosa cells and could be altered in polycystic ovary syndrome (PCOS) from hyperinsulinemia. To determine the relationship between granulosa cell IR isoform expression and follicular fluid insulin concentration in individual follicles, 18 normal women and seven PCOS patients receiving gonadotropins for in vitro fertilization were studied. Glucose tolerance testing was performed before pituitary desensitization, and fasting serum insulin was measured at oocyte retrieval. Granulosa cells and fluid aspirated from the first follicle were used to determine IR isoform mRNA expression and insulin concentration, respectively. IR isoform A mRNA expression was greater than that of IR isoform B expression in normal mural granulosa and cumulus cells, without a cell type effect. Intrafollicular insulin levels increased with adiposity and serum insulin levels at oocyte-retrieval but did not predict IR mRNA expression. Total IR mRNA expression, but not intrafollicular insulin levels, was elevated in PCOS patients, whereas intrafollicular insulin levels were increased in women with impaired glucose tolerance. Granulosa cell IR heterogeneity, together with adiposity-dependent intrafollicular insulin availability, introduces a novel mechanism by which insulin may affect granulosa cell function within the follicle.

Asunto(s)

Insulina/metabolismo , Folículo Ovárico/metabolismo , Síndrome del Ovario Poliquístico/metabolismo , Receptor de Insulina/metabolismo , Adulto , Estudios de Casos y Controles , Células Cultivadas , Femenino , Intolerancia a la Glucosa , Células de la Granulosa/metabolismo , Hirsutismo , Humanos , Ovulación , Síndrome del Ovario Poliquístico/patología , Receptor de Insulina/genética

RESUMEN

A prospective randomized trial was performed to compare post-thaw development of murine blastocysts following programmable rate freezing and two methods of vitrification. Frozen 2-cell murine embryos (n = 429) thawed and cultured for 48 h, were randomly allocated by stage of development into four groups: control (not refrozen), programmable rate freezing (PR) in 0.25 ml straws, vitrification in flexible micropipettes by immersion in super-cooled (VSC) liquid nitrogen (LN2), and vitrification in flexible micropipettes by immersion in LN2 (VLN). Survival, developmental stage progression, presence or absence of an inner cell mass (ICM), and cell counts were recorded 24 h post-thaw. All measured outcomes were different between embryos from the control group and all freezing methods. Controlled-rate freezing resulted in the lowest total cell counts and fewest embryos with a distinct ICM. A higher percentage of embryos survived 24 h post-thaw, progressed to more advanced developmental stages and had higher total cell counts after VLN compared with PR. Moreover, fewer embryos, frozen by either PR or VSC, contained a detectable ICM compared with VLN. These data demonstrate that vitrification may be a better method for freezing murine blastocysts than PR, and may prove to be a superior method for freezing human blastocysts.

Asunto(s)

Blastocisto/fisiología , Criopreservación , Transferencia de Embrión , Animales , Humanos , Ratones

RESUMEN

OBJECTIVE: To estimate cumulative chance for first live birth after elective pronuclear stage cryopreservation of all embryos due to ovarian hyperresponsiveness. DESIGN: Retrospective analysis with longitudinal follow-up. SETTING: Academic hospital. PATIENT(S): Thirty subjects with elective cryopreservation of all embryos due to ovarian hyperresponsiveness. INTERVENTION(S): Elective cryopreservation of all embryos at the pronuclear stage (n = 30) and subsequent cryopreserved-thawed ET (n = 51). MAIN OUTCOME MEASURE(S): Cumulative chance for first live birth. RESULT(S): Cumulative chance for first live birth was 77% when analyzed by intention to treat and 82% by treatment with ET. Nearly 40% of live births were multiple. CONCLUSION(S): Cumulative first live birth increased with repetitive ET after elective pronuclear stage cryopreservation of all embryos due to ovarian hyperresponsiveness. Multiple births, however, were frequent. In the context of initial ET attempts in young women, transfer of no more than two cryopreserved-thawed embryos is advised.

Asunto(s)

Criopreservación/métodos , Embrión de Mamíferos , Fertilización In Vitro/métodos , Síndrome de Hiperestimulación Ovárica/fisiopatología , Resultado del Embarazo , Adulto , Gonadotropina Coriónica/sangre , Transferencia de Embrión , Estradiol/sangre , Femenino , Estudios de Seguimiento , Humanos , Recién Nacido , Estudios Longitudinales , Oocitos/citología , Embarazo , Estudios Retrospectivos , Factores de Tiempo

RESUMEN

OBJECTIVE: To validate the Cryo Bio System (CBS) straw in our current cryopreservation system before using it in clinical practice. DESIGN: A prospective comparison of blastocyst development rates in 278 murine embryos after refreezing and thawing at the two-cell stage against the standard Instruments-Medicine-Veterinarian (IMV) straw used in our cryopreservation program. SETTING: Private IVF laboratory. PATIENT(S): No human subjects or material was used in this study. INTERVENTION(S): Frozen two-cell murine embryos were thawed and randomized into three treatments [1] refreezing in the CBS straws, [2] refreezing in IMV 0.25-mL straws, and [3] control embryos remaining in culture without refreezing. Embryos were refrozen using identical cryoprotectants and identical programmed controlled-rate freezers. After cryopreservation, straws were held in liquid nitrogen for a brief period before thawing and continued culture. MAIN OUTCOME MEASURE(S): Postthaw murine blastocyst development rate. RESULT(S): When the manufacturer's filling and loading protocol was used for the CBS straw there was no significant difference in the blastocyst development rate between CBS (75.0%) and IMV (76.4%) straws. CONCLUSION(S): The CBS straw may be a viable and potentially safer alternative for cryopreservation of human embryos, particularly for patients with known infections.

Asunto(s)

Blastocisto/fisiología , Criopreservación/métodos , Desarrollo Embrionario y Fetal/fisiología , Animales , Femenino , Masculino , Ratones , Embarazo , Estudios Prospectivos , Distribución Aleatoria

RESUMEN

OBJECTIVE: To evaluate the relationship of body mass index (BMI) to uterine receptivity under conditions of programmed hormonal support and standardized embryo quality. DESIGN: Retrospective cohort study.A tertiary referral center. PATIENTS: Ninety-seven consecutive first-cycle recipients of anonymous oocyte donation. After programmed hormone replacement, recipients had transfer of embryos derived from oocyte donation. Anonymous oocyte donors received ovarian stimulation and underwent transvaginal ultrasound-guided oocyte retrieval. SETTING: A receiver operator characteristic (ROC) curve of implantation versus BMI. Area under the ROC curve was 0.51, 95% confidence interval (CI) 0.41-0.62, suggesting no relationship between BMI and implantation. There was no difference in implantation rates between obese (BMI >or=30) and nonobese (BMI <30) recipients, odds ratio 1.1, 95% CI 0.5-2.4. CONCLUSION(S): Uterine receptivity was unimpaired in women with increased BMI when hormonal support and embryo quality were standardized.

Asunto(s)

Índice de Masa Corporal , Implantación del Embrión , Infertilidad Femenina/fisiopatología , Infertilidad Femenina/terapia , Donación de Oocito , Útero/fisiopatología , Adulto , Estudios de Cohortes , Femenino , Humanos , Infertilidad Femenina/complicaciones , Infertilidad Femenina/patología , Persona de Mediana Edad , Obesidad/complicaciones , Oportunidad Relativa , Curva ROC , Estudios Retrospectivos , Resultado del Tratamiento