RESUMEN
PURPOSE: To evaluate the concordance between polarization microscopy and confocal microscopy techniques in the evaluation of the meiotic spindle of human oocytes matured in vivo. METHODS: Prospective study that evaluated oocytes with the first polar extruded body obtained from infertile women who had undergone ovarian stimulation for intracytoplasmic sperm injection. The oocytes with the first polar extruded body were evaluated by polarization microscopy and were then immediately fixed and stained for microtubule and chromatin evaluation by high-performance confocal microscopy. We determined the correlation of polarization microscopy with confocal microscopy in the detection of meiotic oocyte anomalies, and we also evaluated the percentage of oocytes with a visible and non-visible cell spindle by polarization microscopy and with meiotic normality and abnormalities by confocal microscopy. Confidence intervals, Kappa's index and concordance between the methodologies were calculated, considering immunofluorescence microscopy analysis as the golden-standard for evaluating normal spindle and oocyte chromosome distribution. RESULTS: We observed that 72.7% of metaphase II oocytes with a nonvisible meiotic spindle by polarization microscopy showed no meiotic abnormalities by confocal analysis and 55.6% of metaphase II oocytes with a visible meiotic spindle by polarization microscopy were found to be abnormal oocytes by the confocal analysis. Only 44.4% of oocytes with a visible meiotic spindle by polarization microscopy were found to be normal by confocal analysis. Concordance between the methods was 51.1% (Kappa: 0.11; 95%CI -0.0958 - 0.319). CONCLUSIONS: The low correlation between polarization microscopy and confocal microscopy in the assessment of oocyte meiotic spindle suggests that visualization of the meiotic spindle of human oocytes at metaphase II by polarization microscopy is not a good indicator of oocyte meiotic normality.
Asunto(s)
Oocitos/ultraestructura , Inducción de la Ovulación , Huso Acromático , Femenino , Humanos , Microscopía Confocal , Microscopía de Polarización , Embarazo , Estudios ProspectivosRESUMEN
OBJECTIVE: To compare oocyte maturation, fertilization and cleavage rates, and embryonic developmental quality after culture of human immature oocytes from polycystic ovary syndrome (PCOS) patients in human tubal fluid (HTF) or tissue culture medium (TCM) 199. DESIGN: Prospective, randomized, controlled trial. SETTING: University hospital. PATIENT(S): Thirteen women undergoing 23 in vitro maturation cycles, from whom 119 oocytes were retrieved. INTERVENTION(S): Cumulus-enclosed germinal vesicle-stage oocytes matured in TCM-199-supplemented or HTF-supplemented media. MAIN OUTCOME MEASURE(S): Oocyte maturation and fertilization rates, embryonic developmental quality. RESULT(S): Significant differences were observed between TCM 199 and HTF regarding maturation rate (82% vs. 56.9%), fertilization rate (70% vs. 39.4%), and embryo quality (81.3% vs. 41.7%). CONCLUSION(S): Human tubal fluid medium, although widely used for embryo fertilization and maintenance in IVF techniques, is not an appropriate medium for the maturation of oocytes obtained from PCOS patients in nonstimulated cycles.
Asunto(s)
Técnicas de Cultivo de Célula , Medios de Cultivo , Trompas Uterinas/metabolismo , Infertilidad Femenina/terapia , Recuperación del Oocito , Oocitos , Oogénesis , Síndrome del Ovario Poliquístico/terapia , Adulto , Fase de Segmentación del Huevo , Técnicas de Cultivo de Embriones , Transferencia de Embrión , Femenino , Fertilización In Vitro , Humanos , Infertilidad Femenina/etiología , Infertilidad Femenina/fisiopatología , Síndrome del Ovario Poliquístico/complicaciones , Síndrome del Ovario Poliquístico/fisiopatología , Embarazo , Estudios ProspectivosRESUMEN
PURPOSE: to evaluate the meiotic spindle and the chromosome distribution of in vitro matured oocytes obtained from stimulated cycles of infertile women with polycystic ovary syndrome (PCOS) and with male factor and/or tubal infertility (Control Group) and compare in vitro maturation (IVM) rates between the groups analyzed. METHODS: five infertile patients with PCOS and eight controls, submitted to stimulated cycles for intracytoplasmic sperm injection, were selected prospectively and consecutively, and respectively assigned to the study group and the Control Group. Immature oocytes (21 and 29, respectively, from PCOS and Control Group) were submitted to IVM. After IVM, oocytes with first polar body extruded were fixed and submitted to immunostaining and fluorescence microscopy for morphological evaluation of the spindle and of chromosome distribution. Statistical analysis was performed by the Fisher test with significance, when p< or =0.05. RESULTS: IVM rates were similar between groups (47.6 e 44.8%, respectively, for PCOS and Control Group). Six of the ten oocytes (60%) from the study group and four of the 12 oocytes (33.3%) from the Control Group presented meiotic anomalies of the spindle and/or anomalous chromosome distribution, without statistical difference between groups. CONCLUSIONS: data from the present study did not demonstrate significant difference neither in IVM rates nor in the proportions of meiotic anomalies between in vitro matured oocytes obtained from stimulated cycles from PCOS patients and control ones.