Supplementing single step culture media with insulin for continuous uninterrupted in vitro culture of human embryos and monitoring the outcome: prospective randomized clinical trial

Abstract

Mono media is a single formulation. Mono formulations require less manipulation and are less expensive. Both formulations have attained good outcomes, but an adequately powered assessment including usable blastulation rates (USBR), ploidy risk, and SIR is lacking.DESIGN: Paired RCT. MATERIALS AND METHODS: Patients with normal ovarian reserve were recruited. A paired design allowed each patient to serve as their own control, eliminating many confounding variables seen in prior studies. After confirming fertilization, patients' zygotes were randomized (1:1 ratio) to either Seq media (Quinn's Advantage Cleavage Medium, Sage then Blast Assist, Origio) or Mono media (Continuous Single Culture; Irvine Scientific). Each culture system used separate incubators. Assessed endpoints for all embryos included USBR, blastulation timing (arrest vs day 5 vs 6) and ploidy status. Paired euploid blastocyst transfers, one from each group, were performed. DNA fingerprinting of concepti was used as needed to link each embryo to a definitive outcome. SIR was defined as the presence of a fetal heart beat at 8-9 weeks gestation. Statistical analysis performed via McNemar's Chi Square and Wilcoxon sum rank tests.RESULTS: 186 patients had their 2PN embryos (N¼2257) randomized to each culture system. Seq media had a higher blastulation rate then Mono media (p¼0.001, 55.5% vs. 46.0%). No differences were found in the day of blastulation (p¼0.4063) or in the aneuploidy rate (p¼0.5518). Of the 168 patients who had euploid blastocysts suitable for transfer, 126 completed a paired embryo transfer and 42 had a SET. Amongst the SETs, the SIR was equivalent (p¼1.0). Of the 126 double embryo transfers, 36 patients had one fetal heart beat at discharge, however there was no statistical difference in the likelihood of implantation between groups (p¼0.8642).CONCLUSIONS: This is the first randomized controlled trial to systematically examine paired euploid transfers of sibling zygotes cultured in in Seq vs. Mono media. This study demonstrates that the usable blastocyst rate is greatest after culture in Seq media in comparison to a Mono formulation; however no difference exists in SIR.Supported by: Irvine scientific provided Mono media.