Evaluation of a novel non-invasive preimplantation genetic screening approach

PLoS One. 2018 May 10;13(5):e0197262. doi: 10.1371/journal.pone.0197262. eCollection 2018.

Abstract

Objective: To assess whether embryonic DNA isolated from blastocyst culture conditioned medium (BCCM) combined with blastocoel fluid (BF) could be used for blastocyst stage non-invasive preimplantation genetic testing for chromosomal aneuploidy (non-invasive preimplantation genetic screening, NIPGS).

Patients: 47 embryos from 35 patients undergoing IVF.

Interventions: DNA analysis of combined BCCM plus BF in comparison with trophectoderm (TE) biopsy and/or whole blastocyst (WB)using next generation sequencing (NGS).

Results: Embryonic DNA was successfully amplified in 47/47 NIPGS samples (28 frozen-thawed and 19 fresh culture samples) ranging from 6.3 to 44.0 ng/μl. For frozen-thawed embryos, the concordance rate for whole chromosome copy number per sample was equivalent between NIPGS vs. TE biopsy, NIPGS vs. WB and TE vs. WB samples taken from the same embryo was 87.5%; 96.4% and 91.7% respectively (P>0.05), and the rate of concordance per single chromosome was 99.3%, 99.7% and 99.7%, respectively (P>0.05). In fresh cases (Day 4 to Day 5/6 culture), the concordance rate for whole chromosome copy number per sample between NIPGS vs. TE samples taken from the same embryo was 100%, and the rate of concordance per single chromosome was 98.2% (P>0.05).

Conclusions: A combination of BCCM and BF contains sufficient embryonic DNA for whole genome amplification and accurate aneuploidy screening. Our findings suggest that aneuploidy screening using BCCM combined with BF could potentially serve as a novel NIPGS approach for use in human IVF.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aneuploidy
  • Blastocyst* / metabolism
  • Culture Media, Conditioned*
  • DNA / metabolism
  • Embryo Culture Techniques
  • Fertilization in Vitro
  • Genetic Testing*
  • Humans
  • Preimplantation Diagnosis*
  • Proof of Concept Study
  • Sequence Analysis, DNA

Substances

  • Culture Media, Conditioned
  • DNA

Grants and funding

Ontario Centres of Excellence TalentEdge Fellowship Program (http://www.oce-ontario.org/programs/industry-academic-collaboration/talentedge/talentedge-fellowship-program) supported in part research activities related to this manuscript by grant #22717 awarded to Dr. Svetlana Madjunkova. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.