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Coticchio G.,Biogenesi Reproductive Medicine Center
Human Reproduction | Year: 2016

Oocyte in vitro maturation (IVM) involves the achievement of the process of oocyte maturation in vitro. Clinically, it was introduced several decades ago as a potentially more patient-friendly and less expensive assisted reproductive technology (ART) approach, as immature oocytes collected from mid-antral follicles can be matured in vitro without prior gonadotrophin stimulation. However, IVM oocytes are developmentally less competent compared with oocytes matured in vivo, a fact that has encouraged the use of short FSH priming and/orhCGtriggering in IVM cycles. These alternatives have generated much confusion about the definition and clinical outcome of IVM, also among ART specialists, especially because hCG triggering can support maturation in vivo even in follicles of 10-13 mm in diameter. In a recent manuscript, a team of IVM specialists propose that IVM should include any ARTapproach involving the collection of oocyte from small and intermediate sized follicles even afterhCG or GnRH triggering. It is more than predictable that other scientists and clinicians with an interest in IVM will object to such a definition, believing that semantically and operatively IVM should not be associated with pharmacological interventions aimed at promoting or achieving maturation in vivo, although partially. © The Author 2016. Source


Coticchio G.,Biogenesi Reproductive Medicine Center
Human reproduction (Oxford, England) | Year: 2013

Are morphometric and morphological parameters of the metaphase II (MII) spindle of human oocytes matured in vivo or in vitro predictive of chromosome alignment on the metaphase plate? Morphometric spindle parameters were very comparable between oocytes matured in vivo and in vitro and were unable to predict chromosome alignment, while a flattened shape of both poles was positively associated with chromosome displacement from the metaphase plate. The relationship between MII spindle morphometry and chromosome alignment has only been sporadically investigated in human oocytes. The possible implications of spindle pole morphology are totally unrecognized. Morphometric and morphological analysis of the MII spindle of donated supernumerary human oocytes (N = 93) aimed at investigating possible associations between novel microtubule parameters and chromosome arrangement. MII oocytes from three sources were analysed: (i) stimulated cycles matured in vivo (ivo-MII), (ii) leftover cumulus-free germinal vesicle oocytes from stimulated cycles matured in vitro (lgv-MII) and (iii) immature cumulus-cell oocyte complexes (COCs) recovered from in vitro maturation (IVM) cycles and matured in vitro (ivm-MII). Oocytes were fixed and stained for tubulin, chromatin and actin. Optical sections were collected at 0.3 μm intervals by high-performance confocal microscopy and three-dimensionally reconstructed for assignment of specific spindle and chromosomal properties. Spindle pole morphology was classified as either focused or flattened depending on whether microtubule ends were more or less convergent, respectively. Optical density measurements were generated to estimate microtubule abundance in chromosome to pole domains proximal and distal to the oolemma. In ivo-MII oocytes, the sizes (mean ± SD) of major and minor axes were 11.8 ± 2.6 and 8.9 ± 1.7 μm, respectively, while maximum projection was 88.8 ± 29.5 μm(2). Very comparable values of these parameters were found in lgv-MII and ivm-MII oocytes. Double-focused spindles were rarely found (3.1%), unlike those with a double-flattened conformation (47.7%). Spindles with both focused and flattened poles amounted to almost half of the sample set (49.2%), but in this subgroup it was very infrequent (4.6%) to observe the flattened pole oriented towards the oolemma. Overall, differences in the relative proportions of pole morphology categories in ivo-MII, lgv-MII and ivm-MII oocytes were not statistically significant. For both the distal and proximal spindle hemidomains, optical intensity profiles were also comparable between ivo-MII, lgv-MII and ivm-MII oocytes. None of the morphometric parameters (major and minor axes, their ratio, maximum projection, distances of the metaphase plate from the poles) was associated with chromosome alignment on the metaphase plate or arrangement inside and outside the spindle. Importantly, a double-flattened outline of pole morphology was positively associated with the displacement of one or more chromosomes from the metaphase plate. Moreover, when a flattened pole was oriented towards the oolemma, a higher rate of chromosome displacement was observed. The findings of the study will require confirmation by further in-depth analysis and extension of the database, especially regarding the relationship between microtubule abundance and chromosome arrangement. Furthermore, considering the high number of comparisons, the observed statistical differences will require future 'ad hoc' analysis. Collectively, this work provides a robust database for future research on the human oocyte cytoskeleton, and contributes to a better definition of oocyte quality in assisted reproduction technology. Also, these data support the notion that IVM does not affect spindle morphometry and morphology. Part of this work was supported by a grant awarded by the Italian Ministry of Labour, Health and Social Policies. The authors have no conflicts of interest to declare. Not applicable. Source


Barberi M.,University of Rome La Sapienza | Di Paolo V.,University of Rome La Sapienza | Latini S.,University of Rome La Sapienza | Guglielmo M.C.,University of Rome La Sapienza | And 3 more authors.
Molecular and Cellular Endocrinology | Year: 2013

Pituitary adenylate cyclase-activating polypeptide (PACAP) and its receptor PAC1-R (PACAP type 1 receptor) are transiently expressed in granulosa cells (GCs) of mouse preovulatory follicles and affect several parameters associated with the ovulatory process. We investigated the expression of PACAP and its receptors in cumulus cells (CCs) after the LH surge and their role on cumulus expansion/apoptosis and oocyte maturation. PACAP and PAC1-R expression increased in CCs isolated at different times after treatment with human chorionic gonadotropin (hCG). Moreover, PACAP was able to reverse the inhibition of oocyte meiotic maturation caused by hypoxantine in cumulus cell-oocyte complexes (COCs) and efficiently promoted male pronuclear formation after fertilisation. PACAP was also able to induce cumulus expansion and prevent CC apoptosis. Our results demonstrated the induction of PACAP and its receptors in CCs by LH and EGF, suggesting that PACAP may play a significant role in the complex interactions of gonadotropin and growth factors during ovulation and fertilisation. © 2013 Elsevier Ireland Ltd. Source


Grondahl M.L.,Copenhagen University | Nielsen M.E.,University of Southern Denmark | Dal Canto M.B.,Biogenesi Reproductive Medicine Center | Fadini R.,Biogenesi Reproductive Medicine Center | And 4 more authors.
Reproductive BioMedicine Online | Year: 2011

This study evaluated whether anti-Müllerian hormone (AMH) was differentially expressed in cumulus (CC) and granulosa (GC) cells from large antral and pre-ovulatory follicles collected from individual follicles in women undergoing in-vitro maturation (IVM) or IVF treatment. Expression studies of AMH, AMH receptor 2, FSH receptor, aromatase and androgen receptor were performed in CC in IVM patients where cumulus-oocyte-complex had expanded, CC in IVM patients where cumulus-oocyte-complex remained compacted, GC from immature follicles and CC and GC from IVF patients. Microarray data on corresponding GC and CC from follicles from IVF patients was included. AMH expression was significantly higher in CC than in GC from both mature and immature follicles and in CC from immature follicles than in CC from pre-ovulatory follicles from IVF patients (P < 0.05). AMH expression was significantly higher in CC that remained compacted compared with those that had expanded (P < 0.008). AMH was correlated to the expression of FSH receptor, androgen receptor and AMH receptor 2 but not to aromatase expression. The expression pattern of AMH receptor 2 reflected that of AMH. AMH may exert intrafollicular functions even in human large antral and pre-ovulatory follicles and may be related to follicular health. © 2010, Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved. Source


Fadini R.,Biogenesi Reproductive Medicine Center | Mignini Renzini M.,Biogenesi Reproductive Medicine Center | Guarnieri T.,Biogenesi Reproductive Medicine Center | Dal Canto M.,Biogenesi Reproductive Medicine Center | And 5 more authors.
Human Reproduction | Year: 2012

STUDY QUESTIONAre the obstetric and perinatal outcomes of deliveries following in vitro maturation (IVM) cycles different from births generated from controlled ovarian stimulation (COS) cycles?SUMMARY ANSWERThe obstetric and perinatal outcomes of births from IVM cycles are comparable with those of ICSI treatments, including the incidence of major and minor abnormalities. WHAT IS KNOWN ALREADYOnly few and numerically small reports on the health of IVM children are currently available. STUDY DESIGN, SIZE AND DURATIONRetrospective cohort study involving 196 babies born from IVM cycles carried out with different priming regimens. Of these children, 79 developed from oocytes matured in vitro after 30 h of culture, while 104 originated from oocytes found mature and inseminated on the day of recovery. Thirteen babies were obtained from embryos developed from both types of oocytes. Data of these births were compared with those of 194 children born from COS ICSI cycles performed during the same period (March 2004 to December 2011). PARTICIPANTS/MATERIALS, SETTING AND METHODSIVM cycles were done in the absence of gonadotrophin administration or with FSH and/or HCG priming. All oocytes were inseminated by microinjection. ICSI and ICSI cycles were chosen as a control group to exclude possible influences of the insemination technique. Couples in which maternal age was >39 years or affected by azoospermia were excluded to rule out major parental effects. MAIN RESULTS AND THE ROLE OF CHANCEIn single births, gestational age at delivery was comparable, but birthweight was significantly higher (P=0. 009) in children from IVM cycles (3091 ± 669 versus 3269 ± 619 g). In a separate analysis of the IVM group, comparing singleton births derived with certainty from oocytes matured in vitro (n=71) or in vivo (n=74), no statistically significant differences were observed in terms of birthweight (3311 ± 637 versus 3194 ± 574 g, respectively) and gestational age (38. 9 ± 2. 4 versus 38. 4 ± 2. 1 weeks, respectively). In twin births, gestational age was lower in IVM cycles, while weight at birth was comparable (ICSI, 2432 ± 540 g; IVM, 2311 ± 577 g). In single births, major and minor abnormalities were 2 (1. 4) and 6 (4. 1) in the ICSI group and 0 (0. 0) and 8 (5. 2) in the IVM category, respectively. In twin children, major and minor abnormalities were 1 (2. 2) and 2 (4. 3) in ICSI babies and 0 (0. 0) and 2 (4. 6) in IVM cycles, respectively. LIMITATIONS AND REASONS FOR CAUTIONThe study is the largest conducted so far. Nevertheless, it is limited by its retrospective nature and the fact that most births of IVM treatments derived from oocytes found mature at recovery in cycles primed with HCG. A more comprehensive appraisal of the health status of IVM children will demand larger prospective studies. WIDER IMPLICATIONS OF THE FINDINGSThe study is consistent with previous reports suggesting a possible role of standard ovarian stimulation in determining a reduced birthweight in children born from COS cycles. STUDY FUNDING/COMPETING INTEREST(S)No external funding was sought to support this work. The authors have no conflicts of interest to declare. TRIAL REGISTRATION NUMBERNot applicable. © 2012 The Author. Source

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