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IOGENESI Centro di Medicina della Riproduzione B OOCYTE SELECTION BASED ON MORPHOLOGICAL CRITERIA AND POLOSCOPE DAL CANTO MARIABEATRICE BIOGENESI REPRODUCTIVE.

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Presentation on theme: "IOGENESI Centro di Medicina della Riproduzione B OOCYTE SELECTION BASED ON MORPHOLOGICAL CRITERIA AND POLOSCOPE DAL CANTO MARIABEATRICE BIOGENESI REPRODUCTIVE."— Presentation transcript:

1 IOGENESI Centro di Medicina della Riproduzione B OOCYTE SELECTION BASED ON MORPHOLOGICAL CRITERIA AND POLOSCOPE DAL CANTO MARIABEATRICE BIOGENESI REPRODUCTIVE MEDICINE CENTER, ISTITUTI CLINICI ZUCCHI V. Zucchi, 24-Monza (Italy) tel/fax

2 IOGENESI Centro di Medicina della Riproduzione B Performing a right evaluation of nuclear and cytoplasmic maturation of human oocytes is extremely important for the result of In Vitro Fertilization: low quality oocytes are hardly going to fertilize or won’t be competent to produce good embryos. More than half of the collected oocytes during IVF show at least one morphologic anomaly that could negatively influence the embryo development.

3 IOGENESI Centro di Medicina della Riproduzione B Morphologic evaluation of oocytes with cumulus-corona complex It’s difficult! It’s necessary “the spreading” of the cumulus-complex (Veeck)

4 IOGENESI Centro di Medicina della Riproduzione B Metaphase II Oocytes without cumulus can be accurately evaluated analyzing the nuclear maturity, the characteristics of the cytoplasm, the aspect of the polar body, the perivitelline space and the ZP.

5 IOGENESI Centro di Medicina della Riproduzione B Two processes need to be synchronize to produce a competent oocyte to use in IVF: Nuclear Maturation can be verified analyzing the presence of the first polar body and the Meiotic spindle signal Cytoplasmic Maturation can be only hypnotized using several morphological criteria based on cytoplasm characteristic

6 IOGENESI Centro di Medicina della Riproduzione B Polar body The first polar body is the real signal of nuclear maturation: Oocyte with 1°polar body is arrested at Metaphase II The first polar body in human remains intact more than 20 hr after ovulation, while in mammals it has a shorter life. We can postulate that the first polar body morphology gives informations not only on the nuclear maturation of the oocytes but on the “AGE” of the oocyte also.

7 IOGENESI Centro di Medicina della Riproduzione B More than a half of the collected oocytes show at least one anomaly. Ebner suggests to divided these anomalies in intracytoplasmic and extracytoplasmic (Ebner T. 2001) Cytoplasmic Maturation

8 IOGENESI Centro di Medicina della Riproduzione B Cytoplasmic Maturation Intracytoplasmic anomalies: variations in colour or granularity of the cytoplasm presence of inclusions, vacuoles or retractable bodies (Ebner et al. 2003, 2005) aggregations of the smooth endoplasmic reticulum (Otsuki J. 2004)

9 IOGENESI Centro di Medicina della Riproduzione B Cytoplasmic Maturation Extracytoplasmic anomalies: include irregularities in the shape of the oocyte, presence of detritus or increment in the perivitelline space,, abnormal consistency of ZP and fragmentation of the first polar body

10 IOGENESI Centro di Medicina della Riproduzione B Oocyte morphology can impact IVF outcome?

11 IOGENESI Centro di Medicina della Riproduzione B Alikani’s group (Alikani 1995) did not find any reduction in the fertilization rate after ICSI on dysmorphic oocytes versus to normal oocytes

12 IOGENESI Centro di Medicina della Riproduzione B Some groups (Balaban et al., 1998; De Sutter et al., 1996) have not found correlation between the oocytes’ morphology and the fertilization rate or the embryo quality after ICSI.

13 IOGENESI Centro di Medicina della Riproduzione B the Serhal’s group (Serhal et al., 1997) and Loutradis’ group (Loutradis et al., 1999), reported higher pregnancy rate in patients in which had been transferred embryos that derived exclusively from oocytes without anomalies

14 IOGENESI Centro di Medicina della Riproduzione B Xia et al. in 1997 correleted the first polar body apparence and periviteline space size with the oocyte’s potenziality to fertilize. Ebner et al. correlated the cytoplasm and polar body morphology with embryo quality and embryo capacity to reach blastocyst stage (Ebner 1999, Ebner 2001, Ebner 2005)

15 IOGENESI Centro di Medicina della Riproduzione B Grade 1° PB N° oocytes % 2PN <25% fragm a 89.0 d b 56.3 e c 47.3 f Influence of first polar body morphology a,b,c p< d,e,f p < Ebner et al The post-maturity of the oocytes seems to be associated to the polar body degeneration It has been found that an integral first polar body is a favourable prognostic factor in terms of fertilization, embryo quality and pregnancy rate.

16 IOGENESI Centro di Medicina della Riproduzione B On the contrary Ciotti (Ciotti et al. 2004) published results against a predictive value of PBI morphology, confirmed by De Santis (De Santis et al. 2005) on fertilization rate and embryo quality. Influence of first polar body morphology De Santis L. 2005

17 IOGENESI Centro di Medicina della Riproduzione B At the stage of metaphase II (second meiotic division) the signal of chromosomes aligned at the equator of the meiotic spindle (MS) can be capture preserving the oocyte’s funzionality. The presence of spindle-signal could be consider a prognostic factor of good quality. The introduction of polarization light microscopy favoured a new approch to analize the oocyte (Wang et al. 2001).

18 IOGENESI Centro di Medicina della Riproduzione B The signal is produced by microtubules located at the oocyte periphery and it is very dynamic structure (Rienzi et al. 2003).

19 IOGENESI Centro di Medicina della Riproduzione B The percentage of oocytes showing a detectable MS varied between 60% and 90%. This difference seems to be related to two important parameters: the high sensitiveness to temperature and the technique of spindle visualization (Wang et al. 2002, Rienzi et al.2003; Cooke et al. 2003).

20 IOGENESI Centro di Medicina della Riproduzione B The position of the signal could be correlate with the “age of oocyte”. Some oocytes showing a first polar body may not have finished nuclear maturation and could be at telophase I stage.

21 IOGENESI Centro di Medicina della Riproduzione B Sometimes oocytes show the spindle far from the polar body. 5% of oocytes show an angle of 45° or 90° between polar body and meiotic spindle (Moon et al., 2003; Rienzi et al., 2003, Rienzi et al.2005)

22 IOGENESI Centro di Medicina della Riproduzione B Rienzi et al. reported that oocytes with visible meiotic spindle have higher fertilization rate than oocytes where it isn’t observable, and the number of high quality embryos and blastocysts increase ( Rienzi et al. 2003) There are no difference in fertilization rate and embryo quality if the angle of displacement does not exceed 90°; if the angle is >90°, lower fertilization rate is observed. (Moon et al., 2003; Rienzi et al., 2003, Rienzi et al.2005) Oocyte at telophase I stage failed the fertilization process. (De Santis et al. 2005)

23 IOGENESI Centro di Medicina della Riproduzione B According to the Italian law we cannot select embryos, we cannot use more than three oocytes per cycle. Since April 2005 we’ve started to select oocytes before ICSI in order to obtain the best fertilization and high quality embryos to transfer. Our STRICT criteria have been: Meiotic spindle: present absent or irregular (45°, 90°) Polar body: regular anomalous Cytoplasmic: regular anomalous (intra+extra anomalies) Our study

24 IOGENESI Centro di Medicina della Riproduzione B Our study 2286 oocytes regularanomalous Spindle % % Polar body % % Cytoplasm % %

25 IOGENESI Centro di Medicina della Riproduzione B Oocyte morphology and fertilization

26 IOGENESI Centro di Medicina della Riproduzione B 1 PN or 3 PN % % degenerated % % p< Meiotic spindle 2286visiblenot visible n° MII Inseminated No fertilization % % 2 PN % %

27 IOGENESI Centro di Medicina della Riproduzione B P= ns 1 PN or 3 PN % % degenerated % % Polar Body 2285regularirregular n° MII Inseminated No fertilization % % 2 PN % %

28 IOGENESI Centro di Medicina della Riproduzione B P< PN or 3 PN % % degenerated % % Cytoplasm 2285regularirregular n° MII Inseminated No fertilization % % 2 PN % %

29 IOGENESI Centro di Medicina della Riproduzione B ParameterMultiple linear regression Fertilization rate Coefficient (95% CI)p-value Meiotic Spindle Polar Body Cytoplasm 1.61 ( ) 0.93 ( ) 1.42 ( ) < < Logistic Regression Analysis : Meiotic spindle and cytoplasm morphology play a SIGNIFICANT INDIPENDENT role on Fertilization

30 IOGENESI Centro di Medicina della Riproduzione B Oocyte morphology and embryo quality

31 IOGENESI Centro di Medicina della Riproduzione B Embryo quality (% of fragmentation) MS +- 0% % % 10% % % 11-20% % % 21-30% % % >30% % % PB % % % % % % % % % % Results P= ns CYTO % % % % % % % % % %

32 IOGENESI Centro di Medicina della Riproduzione B Results 1° step 2° step ?

33 IOGENESI Centro di Medicina della Riproduzione B Results 1° step 2° step 4° step 3° step Women ???

34 IOGENESI Centro di Medicina della Riproduzione B Thank you very much for you attention


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