Dissertation committee:
Δημήτριος Λουτράδης, Καθηγητής, Ιατρική, ΕΚΠΑ
Μιχαήλ Κουτσιλιέρης, Καθηγητής, Ιατρική, ΕΚΠΑ
Παναγούλα Αγγελογιάννη, Επίκουρη Καθηγήτρια, Ιατρική, ΕΚΠΑ
Κωνσταντίνος Καλλιανίδης, Αναπληρωτής Καθηγητής, Ιατρική, ΕΚΠΑ
Νικόλαος Βλάχος, Καθηγητής, Ιατρική, ΕΚΠΑ
Πέτρος Δρακάκης, Αναπληρωτής Καθηγητής, Ιατρική, ΕΚΠΑ
Σταυρούλα Μιχαλά, Επίκουρος Καθηγήτρια, Ιατρική, ΕΚΠΑ
Summary:
Purpose 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, embryo 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 The exposure of human unfertilized oocytes 18 h after ICSI to a combination of calcium ionophore A23187 and the GM-CSF cytokine can safely salvage the oocytes, resulting in the development of good-morphology, euploid blastocysts, with potential clinical use.
Keywords:
Calcium ionophore A23187, GM-CSF, Artificial oocyte activation, ICSI, Fertilization failure
