Treatment with Ca2+ ionophore improves embryo development and outcome in cases with previous developmental problems: a prospective multicenter study.
OBJECTIVE
Does calcium ionophore treatment (A23187, calcimycin) improve embryo development and outcome in patients with a history of developmental problems/arrest?


CONCLUSIONS
Application of A23187 leads to increased rates of cleavage to 2-cell stage, blastocyst formation and clinical pregnancy/live birth.


BACKGROUND
Studies on lower animals indicate that changes in intracellular free calcium trigger and regulate the events of cell division. In humans, calcium fluctuations were detected with a peak shortly before cell division. Interestingly, these calcium oscillations disappeared in arrested embryos. Mitotic division blocked with a Ca(2+) chelator could be restored by means of ionophores in an animal model.


METHODS
This prospective, multicenter (five Austrian centers), uncontrolled intervention study (duration 1 year) includes 57 patients who provided informed consent.


METHODS
Inclusion criteria were complete embryo developmental arrest in a previous cycle (no transfer), complete developmental delay (no morula/blastocyst on Day 5), or reduced blastocyst formation on Day 5 (≤15%). Severe male factor patients and patients with <30% fertilization rate after ICSI were excluded because these would be routine indications for ionophore usage. The total of the 57 immediately preceding cycles in the same patients constituted the control cycles/control group. In the treatment cycles, all metaphase II-oocytes were exposed to a commercially available ready-to-use ionophore for 15 min immediately after ICSI. After a three-step washing procedure, in vitro culture was performed as in the control cycles, up to blastocyst stage when achievable.


RESULTS
Fertilization rate did not differ (75.4 versus 73.2%); however, further cleavage to 2-cell stage was significantly higher (P < 0.001) in the ionophore group (98.5%) when compared with the control cycles (91.9%). In addition, significantly more (P < 0.05) blastocysts formed on Day 5 in the study compared with the control group (47.6 versus 5.5%, respectively) and this was associated with a significant increase (P < 0.01) in the rates of implantation (44.4 versus 12.5%), clinical pregnancy (45.1 versus 12.8%) and live birth (45.1 versus 12.8%). All babies born at the time of writing (22/28) were healthy.


CONCLUSIONS
The frequency of patients showing embryo developmental problems was expected to be low; therefore, a multicenter approach was chosen in order to increase sample size. In one-third of the cycles, the clinician or patient requested a change of stimulation protocol; however, this did not influence the developmental rate of embryos.


CONCLUSIONS
This is the first evidence that developmental incompetence of embryos is an additional indication for ionophore treatment. The present approach is exclusively for overcoming cleavage arrest.


BACKGROUND
No funding received. T.E. reports fees from Gynemed, outside the submitted work. All co-authors have no interest to declare.
Department of Gynecological Endocrinology and Kinderwunsch Zentrum, Landes-Frauen-und Kinderklinik, Krankenhausstr. 26-30, A-4020 Linz, Austria Faculty of Medicine, Johannes Kepler University, 4020 Linz, Austria thomas.ebner@gespag.at.
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Ebner
Department of Gynecological Endocrinology and Kinderwunsch Zentrum, Landes-Frauen-und Kinderklinik, Krankenhausstr. 26-30, A-4020 Linz, Austria Faculty of Medicine, Johannes Kepler University, 4020 Linz, Austria.
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Oppelt
Kinderwunschzentrum Goldenes Kreuz, 1090 Vienna, Austria.
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Wöber
Medical University, 8010 Graz, Austria.
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Staples
Department of Gynecological Endocrinology and Kinderwunsch Zentrum, Landes-Frauen-und Kinderklinik, Krankenhausstr. 26-30, A-4020 Linz, Austria Faculty of Medicine, Johannes Kepler University, 4020 Linz, Austria.
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Mayer
Babywunsch-Klinik Dr. Zajc, 5020 Salzburg, Austria.
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Sonnleitner
Sterignost, Institut für Kinderwunschbehandlung, 9020 Klagenfurt, Austria.
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Bulfon-Vogl
Kinderwunschambulanz im Landesklinikum St Pölten, 3100 St Pölten, Austria.
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Gruber
Department of Gynecological Endocrinology and Kinderwunsch Zentrum, Landes-Frauen-und Kinderklinik, Krankenhausstr. 26-30, A-4020 Linz, Austria Faculty of Medicine, Johannes Kepler University, 4020 Linz, Austria.
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Haid
Department of Gynecological Endocrinology and Kinderwunsch Zentrum, Landes-Frauen-und Kinderklinik, Krankenhausstr. 26-30, A-4020 Linz, Austria Faculty of Medicine, Johannes Kepler University, 4020 Linz, Austria.
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Shebl
D002118:Calcium / Q000378:metabolism; D061207:Calcium Ionophores / Q000494:pharmacology*; D046149:Embryo Culture Techniques; D004624:Embryo Transfer; D047108:Embryonic Development / Q000187:drug effects*; D006801:Humans; D011446:Prospective Studies
D061207:Calcium Ionophores; D002118:Calcium
blastocyst formation; calcium ionophore; cleavage to 2-cell stage; developmental arrest; developmental delay