Busting the myth of extended blastocyst culture until Day 7: Protocol for systematic review and meta-analysis

BACKGROUND: The prolonged culture of embryos to the blastocyst stage represents an increasing procedure in Assisted Reproductive Technology (ART) laboratories. Generally, only blastocysts developing on Day 5 and Day 6 are considered suitable embryos for transfer, cryopreservation or biopsy while embryos developed at a slower rate after Day 6 are routinely discarded. However, also blastocysts developing on Day 7 can be viable and result in a healthy live birth. Unfortunately, data regarding the clinical outcomes of Day 7 blastocysts compared to blastocysts developing on Day 5 or Day 6 are controversial. In this systematic review and aggregate data meta-analysis, we aim to evaluate the real reproductive potential of delayed blastocysts on Day 7 in frozen cycles. METHODS: We will include all studies, with no restriction regarding the study design (randomized and observational trials, including cohort and case-control), investigating the clinical success of blastocysts developed on Day 7 compared to Day 5 and Day 6 blastocysts. The primary outcomes are the clinical pregnancy rate (CPR) and ongoing pregnancy rate (OPR) following frozen-thawed embryo transfer (Day 7 vs Day 6, and Day 5); secondary outcomes are: live birth rate (LBR) following frozen-thawed embryo transfer, euploid rate and survival rate of thawed blastocyst. Two reviewers independently will judge the methodological quality of studies included in the meta-analysis using the criteria reported in the Cochrane Handbook for Systematic Reviews of Interventions or the Newcastle-Ottawa Scale according to the design of the trials. The meta-analysis will be performed using random effects models and heterogeneity will be assessed using Higgins I2 value. Summary estimate of the proportion of each outcome will be expressed as pooled proportion with 95% confidence interval (CI). The effect of the day on each outcome will be evaluated using a multilevel mixed-effects model with a moderator (the day). The effect will be expressed as odds ratio (OR) with 95% confidence interval (CI). A P value less than .05 will be considered statistically significant. ETHICS AND DISSEMINATION: This is a systematic review not requiring an ethical approval. Findings derived from this systematic review and meta-analysis will be published in a peer-reviewed journal.