title: The combination of calcium ionophore A23187 and GM-CSF can safely salvage aged human unfertilized oocytes after ICSI.
abstract: OBJECTIVE
  Artificial oocyte activation using calcium ionophores and enhancement of embryonic developmental potential by the granulocyte-macrophage colony-stimulating factor (GM-CSF) have already been reported. In this study, we evaluated the synergistic effect of these two methods on aged human unfertilized oocytes after intracytoplasmic sperm injection (ICSI). Then, we cultured the resulting embryos to the blastocyst stage and screened them for chromosomal abnormalities, to assess the safety of this protocol.
  
  
  METHODS
  Aged human oocytes deemed unfertilized after ICSI were activated, either by briefly applying the calcium ionophore A23187 alone (group A) or by briefly applying the ionophore and then supplementing the culture medium with recombinant human GM-CSF (rhGM-CSF) (group B). Next, the development was monitored in a time-lapse incubator system, and ploidy was analyzed by array comparative genomic hybridization (aCGH), after whole embryo biopsy and whole genome amplification. Differences between oocytes and resulting embryos in both groups were evaluated statistically.
  
  
  RESULTS
  Oocytes unfertilized after ICSI can be activated with the calcium ionophore A23187 to show two pronuclei and two polar bodies. Addition of rhGM-CSF in the culture medium of A23187-activated oocytes enhances their cleaving and blastulation potential and results in more euploid blastocysts compared to the culture medium alone.
  
  
  CONCLUSIONS
  This study shows that activating post-ICSI aged human unfertilized oocytes with a combination of a calcium ionophore and a cytokine can produce good-morphology euploid blastocysts.
authors.affiliation: Embryogenesis, Assisted Reproduction Unit, 49 Kifissias Avenue and Ziridi Street, 151 23 Maroussi, Athens, Greece. keconomou@gmail.com.
authors.forename: Konstantinos A
authors.initials: KA
authors.lastname: Economou
authors.affiliation: Embryogenesis, Assisted Reproduction Unit, 49 Kifissias Avenue and Ziridi Street, 151 23 Maroussi, Athens, Greece.
authors.forename: Dimitra
authors.initials: D
authors.lastname: Christopikou
authors.affiliation: Embryogenesis, Assisted Reproduction Unit, 49 Kifissias Avenue and Ziridi Street, 151 23 Maroussi, Athens, Greece.
authors.forename: Erika
authors.initials: E
authors.lastname: Tsorva
authors.affiliation: Embryogenesis, Assisted Reproduction Unit, 49 Kifissias Avenue and Ziridi Street, 151 23 Maroussi, Athens, Greece.
authors.forename: Stephen
authors.initials: S
authors.lastname: Davies
authors.affiliation: Embryogenesis, Assisted Reproduction Unit, 49 Kifissias Avenue and Ziridi Street, 151 23 Maroussi, Athens, Greece.
authors.forename: Minas
authors.initials: M
authors.lastname: Mastrominas
authors.affiliation: Embryogenesis, Assisted Reproduction Unit, 49 Kifissias Avenue and Ziridi Street, 151 23 Maroussi, Athens, Greece.
authors.forename: Haris
authors.initials: H
authors.lastname: Cazlaris
authors.affiliation: Department of Physiology, School of Medicine, National and Kapodistrian University of Athens, Athens, Greece.
authors.forename: Michael
authors.initials: M
authors.lastname: Koutsilieris
authors.affiliation: Department of Physiology, School of Medicine, National and Kapodistrian University of Athens, Athens, Greece.
authors.forename: Panagoula
authors.initials: P
authors.lastname: Angelogianni
authors.affiliation: First Department of Obstetrics and Gynaecology, School of Medicine, Alexandra University Hospital, National and Kapodistrian University of Athens, Athens, Greece.
authors.forename: Dimitris
authors.initials: D
authors.lastname: Loutradis
mesh_terms: D001755:Blastocyst / Q000187:drug effects; D000001:Calcimycin / Q000008:administration & dosage; D061207:Calcium Ionophores / Q000008:administration & dosage; D055028:Comparative Genomic Hybridization; D003470:Culture Media / Q000737:chemistry; D047108:Embryonic Development / Q000187:drug effects* / Q000235:genetics; D005260:Female; D005307:Fertilization in Vitro; D006801:Humans; D008297:Male; D009865:Oocytes / Q000187:drug effects* / Q000254:growth & development; D020554:Sperm Injections, Intracytoplasmic
chemical_list: D061207:Calcium Ionophores; D003470:Culture Media; D000001:Calcimycin
keywords: Artificial oocyte activation; Calcium ionophore A23187; Fertilization failure; GM-CSF; ICSI; aCGH