L. Escrich

Morphologic indicators predict the stage of chromatin condensation of human germinal vesicle oocytes recovered from stimulated cycles

OBJECTIVE To assess germinal vesicles (GV) recovered from stimulated cycles by means of morphometric and morphologic examination (using contrast-phase and image analysis) and chromatin configuration (using fluorescent DNA imaging), and to evaluate the relevance of morphometric and morphologic parameters as forecasters of chromatin status. DESIGN Experimental study. SETTING University-affiliated infertility clinic. PATIENT(S) One hundred and thirty-one GV oocytes donated to patients for intracytoplasmic sperm injection. INTERVENTION(S) We evaluated 131 GVs by means of morphology and morphometry with the use of contrast phase microscopy. They were subsequently fixed, DNA stained, and assessed by fluorescent microscopy. Compiled data were retrospectively grouped according to three models. MAIN OUTCOME MEASURE(S) Model A: ova were grouped according to chromatin condensation (noncondensed vs. condensed). Model B: ova were grouped according to chromatin distribution in relation to the nucleolus-like body (NLB) (not surrounding vs. surrounding and/or absent) but regardless of the condensation stage. Model C: GV oocytes were grouped according to the combination of both of the previously mentioned parameters (chromatin condensation and distribution in relation to the NLB). RESULT(S) According to the GV classification of model A, nucleoplasm, nucleus position, nuclear envelope continuity, and oocyte size were shown to be relevant and were included in a mathematical model for predicting chromatin condensation stage. CONCLUSION(S) Noninvasive analysis of GV oocytes using contrast-phase microscopy maintains oocytes in a viable state and allows the chromatin condensation status to be predicted.

The dynamics of in vitro maturation of germinal vesicle oocytes

OBJECTIVE To evaluate the dynamics of the nuclear maturation (NM) of in vitro-matured (IVM) oocytes and to determine the most favorable duration of meiosis II (MII) arrest in relation to the normal activation response. DESIGN Experimental. SETTING University-affiliated infertility clinic. PATIENT(S) Donated immature germinal vesicle oocytes (GV). INTERVENTION(S) The GV underwent spontaneous IVM and the dynamics of NM studied by real-time monitoring. The IVM oocytes were parthenogenetically activated at different MII arrest points and their response assessed. MAIN OUTCOME MEASURE(S) Moment of GV breakdown; extrusion of the first polar body; duration of MI and MII arrest; activation rate (AR) and type. RESULT(S) Two GV populations-early (E-IVM, 18.4 ± 2.7 hours) and late (L-IVM, 26.3 ± 3.8 hours) maturing-were defined according to the time required for extrusion of the first polar body. Significantly more E-IVM than L-IVM exhibited a normal activation response (61.3% vs. 34.6%), but AR were similar (average, 88.6%) in both groups. Duration of the GV stage differed between the two groups, but MI arrest (14.0 ± 0.3 hours) was constant. The E-IVM arrested at MII for at least 4.3 hours displayed significantly lower AR and similar normal activation rates (61.3%) to E-IVM arrested for a shorter time (83.9% vs. 100%). The L-IVM displayed a similar AR (80.8%), but lower normal activation rates than E-IVM (34.6%), regardless of when activation took place. CONCLUSION(S) The success of IVM depends on the NM timing rather than on the length of MII arrest.

Vitrification of isolated human blastomeres

This article describes a new methodology for preserving and banking isolated human blastomeres, whose originality is based on packing the blastomere into an emptied zona pellucida before vitrification. After warming, 75.7% of blastomeres survived and developed at a rate comparable to that in noncryopreserved blastomeres (62.5% cleavage, 26.6% compaction, and 20.3% cavitation).

Do immature and mature sibling oocytes recovered from stimulated cycles have the same reproductive potential

RESEARCH QUESTION How can laboratory and clinical outcomes of spontaneously, early maturing germinal-vesicle oocytes and sibling in-vivo-matured (metaphase II [MII]) oocytes be quantified and compared? DESIGN A prospective, non-randomized intra-cohort study of oocytes from women aged 38 years or younger, with six or fewer MII oocytes and four or more germinal vesicles retrieved. No indication was identified for genetic tests or oocyte or embryo cryopreservation. The study was carried out at IVIRMA-Valencia. Early maturing germinal vesicles were selected for reproductive purposes. In vitro- and in-vivo MII oocytes were fertilized. After time-lapse culture, hatching blastocysts from germinal vesicles were biopsied for aneuploidy screening and vitrified. Laboratory and clinical outcomes were compared according to oocyte origin. RESULTS Almost 70% of germinal vesicles had matured early and spontaneously, and had comparable in vitro-outcomes and morphokinetics to sibling in vivo-matured oocytes. Fifty per cent of biopsied blastocysts were euploid. Germinal-vesicle rescue increased the number of MII oocytes per cycle to 3.9, finally adding one extra-blastocyst per cycle. A live birth confirmed the feasibility of this approach. Further data, however, are needed to quantify its real contribution to standard intracytoplasmic sperm injection cycles. Nevertheless, 40% of patients obtained either an immediate advantage (reduction of cancellation rate) or long-term benefit (availability of extra blastocysts of attempts). CONCLUSIONS Germinal-vesicle rescue can be considered as a complementary approach when folliculometry (expected) and number of MII (observed) are unequal.