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ASSISTED HATCHING & FRAGMENT REMOVAL DO THEY HAVE AND IMPACT IN ART CYCLES Gloria Calderón PhD

ASSISTED HATCHING & FRAGMENT REMOVAL DO THEY HAVE AND IMPACT IN ART CYCLES Gloria Calderón PhD. ASSISTED HATCHING Hypothesis. Zona pellucida anomalies can affect blastocyst hatching “in vivo”. FRAGMENT REMOVAL: Hypothesis. The presence of fragments “per se” can interfere

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ASSISTED HATCHING & FRAGMENT REMOVAL DO THEY HAVE AND IMPACT IN ART CYCLES Gloria Calderón PhD

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  1. ASSISTED HATCHING & FRAGMENT REMOVAL • DO THEY HAVE AND IMPACT IN ART CYCLES • Gloria Calderón PhD

  2. ASSISTED HATCHINGHypothesis • Zona pellucida anomalies can affect blastocyst hatching “in vivo”

  3. FRAGMENT REMOVAL:Hypothesis • The presence of fragments “per se” can interfere • in human embryo development

  4. Presence of fragments & poor embryo morphology • Some mechanism of action proposed: • Active: • Apoptosis induction • Jurisicova et al. Mol Hum Reprod 1996,2:93-98 • Passive • Kidnapping of regulatory proteins • Antczak y Van Blerkom Hum Reprod 1999,14:429-447 • Mechanical interference among blastomeres avoiding their contact • Edwards y Beard Mol Hum Reprod 1997, 3:863-905 • None of this models has been demonstrated

  5. Assisted Hatching: Indications • Zona pellucida anomalies • Embryos with ≤ 6 cells on day 3 of development • Multinucleated embryos • Frozen-thawed embryos • Implantation failure

  6. Embryos with > 10% fragments on day 3 of development Thawed embryos with degenerated blastomeres Fragment removal: Indications

  7. MATERIAL & METHODS • Single culture of embryos in HTF media • Embryo morphology evaluation on D2 & D3 in Olympus IX70 (x600) microscope • % of fragments defined as the occupied volume in the embryo and expressed in %

  8. ASSISTED HATCHING & FRAGMENT REMOVAL • Tyrode’s Acid • No Laser • Micropipettes of 10-12 µm external diameter • Fragment removal

  9. AHA & FRAGMENT REMOVAL

  10. FRAGMENT REMOVAL • Alikani et al Fertil Steril 1999, 71:836-842

  11. RESULTS: % OF FRAGMENTS Alikani et al. 1999

  12. RESULTS: FRAGMENTATION PATTERNS Alikani et al. 1999

  13. Assisted Hatching &Fragment Removal: IVI-SEVILLA

  14. PREVIOUS STUDIES: LEARNING CURVE

  15. PREVIOUS STUDIES: LEARNING CURVE

  16. PREVIOUS STUDIES a, b: p <0,.05 c, d: p <0,.05

  17. MATERIAL & METHODS • Retrospective study of 368 IVF cycles. Only regular patients included. • Assisted hatching performed with tyrode´s acid at least 1h before replacement. • Group I (n=227): all embryos ≤ 10% frag. –AF. • Group II (n=63): at least one embryo >10% frag. –AF. • Group III (n=78): at least one embryo >10% frag. +AF.

  18. a b b RESULTS a, b: p <0,.05

  19. RESULTS: % FRAGMENTS ON D3

  20. c a b d ab c CLINICAL RESULTS a, b: p <0,.05 c, d: p <0,.05

  21. CONCLUSIONS • After fragment removal, embryos with 5-35% of fragments have the same implantation potential. • Fragment removal allows better contact between blastomeres promoting normal compaction and blastocyst formation.

  22. CONCLUSIONS • Pregnancy and implantation rates are affected by the quality of embryos selected for replacement. • The use of Assisted Hatching combined with Fragment removal improves considerably the implantation potential of replaced embryos.

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