title: Proof of concept: preimplantation genetic screening without embryo biopsy through analysis of cell-free DNA in spent embryo culture media.
abstract: OBJECTIVE
  To assess whether preimplantation genetic screening (PGS) is possible by testing for free embryonic DNA in spent IVF media from embryos undergoing trophectoderm biopsy.
  
  
  METHODS
  Prospective cohort analysis.
  
  
  METHODS
  Academic fertility center.
  
  
  METHODS
  Seven patients undergoing IVF and 57 embryos undergoing trophectoderm biopsy for PGS.
  
  
  METHODS
  On day 3 of development, each embryo was placed in a separate media droplet. All biopsied embryos received a PGS result by array comparative genomic hybridization. Preimplantation genetic screening was performed on amplified DNA extracted from media and results were compared with PGS results for the corresponding biopsy.
  
  
  METHODS
  [1] Presence of DNA in spent IVF culture media. [2] Correlation between genetic screening result from spent media and corresponding biopsy.
  
  
  RESULTS
  Fifty-five samples had detectable DNA ranging from 2-642 ng/μL after a 2-hour amplification. Six samples with the highest DNA levels underwent PGS, rendering one result with a derivative log ratio SD (DLRSD) of <0.85 (a quality control metric of oligonucleotide array comparative genomic hybridization). The fluid sample and trophectoderm results were identical demonstrating (45XY, -13). Three samples were reamplified 1 hour later and tested showing improving DLRSD. One of the three samples with a DLRSD of 0.85 demonstrated (46XY), consistent with the biopsy. Overnight DNA amplification showed DNA in all samples.
  
  
  CONCLUSIONS
  We demonstrate two novel findings: the presence of free embryonic DNA in spent media and a result that is consistent with trophectoderm biopsy. Improvements in DNA collection, amplification, and testing may allow for PGS without biopsy in the future.
authors: Fertility and Surgical Associates of California, Thousand Oaks, California; University of California, Los Angeles, Fertility and Reproductive Health Center, Los Angeles, California. Electronic address: mshamonki@gmail.com.
authors: Mousa I
authors: MI
authors: Shamonki
authors: PacGenomics, Agoura Hills, California.
authors: Helen
authors: H
authors: Jin
authors: ART Reproductive Center, Beverly Hills, California.
authors: Zachary
authors: Z
authors: Haimowitz
authors: PacGenomics, Agoura Hills, California.
authors: Lian
authors: L
authors: Liu
mesh_terms: D000046:Academic Medical Centers; D000328:Adult; D001706:Biopsy; D001755:Blastocyst / Q000378:metabolism*; D055028:Comparative Genomic Hybridization; D017077:Culture Media, Conditioned / Q000378:metabolism; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D046149:Embryo Culture Techniques; D005260:Female; D005298:Fertility; D005307:Fertilization in Vitro / Q000009:adverse effects; D005820:Genetic Testing / Q000379:methods*; D006801:Humans; D007246:Infertility / Q000175:diagnosis / Q000503:physiopathology / Q000628:therapy*; D008297:Male; D011237:Predictive Value of Tests; D011247:Pregnancy; D019836:Preimplantation Diagnosis / Q000379:methods*; D011446:Prospective Studies; D015203:Reproducibility of Results
chemical_list: D017077:Culture Media, Conditioned; D004247:DNA
keywords: In vitro fertilization (IVF); comprehensive chromosomal screening (CCS); free embryonic DNA; trophectoderm biopsy (PGS)