AUTHOR=Shi Wenhao , Zhao Zhenghao , Xue Xia , Li Qian , Yao Yaxin , Wang Dongyang , Wang Jing , Lu Sijia , Shi Juanzi 

TITLE=Ploidy Testing of Blastocoel Fluid for Screening May Be Technically Challenging and More Invasive Than That of Spent Cell Culture Media

JOURNAL=Frontiers in Physiology

VOLUME=13

YEAR=2022

URL=https://www.frontiersin.org/journals/physiology/articles/10.3389/fphys.2022.794210

DOI=10.3389/fphys.2022.794210

ISSN=1664-042X

ABSTRACT=<sec><title>Background</title><p>Recent studies have demonstrated that both blastocoel fluid (BF) and spent cell culture media (SCM) have potential as materials for non-invasive or less-invasive pre-implantation genetic analysis. BF may allow more opportunity to obtain cell-free DNA from the inner cell mass (ICM), and it has a lower risk of containing contaminant DNA from cumulus cells, sperm and culture media. There are no data regarding the ICM as a gold standard to evaluate the chromosome constitution of BF or SCM for embryo liquid biopsy.</p></sec><sec><title>Methods</title><p>Two hundred eighteen donated human blastocysts were warmed and cultured in blastocyst culture media for 18–24 h. The corresponding SCM was collected, and only clear ICM was biopsied in blastocysts; otherwise, the whole blastocyst (WB) was biopsied. Quantitative PCR was performed to determine the DNA levels in the SCM and BF before and after amplification. ChromInst was used to amplify BF/SCM and blastocyst DNA before sequencing. Chromosomal copy number variation (CNV) was investigated to evaluate the chromosome constitution.</p></sec><sec><title>Results</title><p>In total, 212 blastocysts were available for SCM and BF collection. The technical success rates (next-generation sequencing data) were 100 and 69.8% (148/212) for SCM and BF, respectively. Among the 148 blastocysts with both SCM and BF data, 101 were euploid and 47 were aneuploid based on ICM (<italic>n</italic> = 89) or WB (<italic>n</italic> = 59) analysis as the gold standard. Among all blastocysts, SCM was comparable to BF [specificity: 80.2 versus 61.4% (<italic>P</italic> = 0.005, χ<sup>2</sup> test); sensitivity: 91.5 versus 87.2% (<italic>P</italic> = 0.738, χ<sup>2</sup> test); negative predictive value (NPV): 95.3 versus 91.2% (<italic>P</italic> = 0.487, χ<sup>2</sup> test); positive predictive value (PPV): 68.3% versus 51.3% (<italic>P</italic> = 0.042, χ<sup>2</sup> test)]. The SCM and BF samples were 83.8% (124/148) and 69.6% (103/148) concordant with the corresponding ICM/WB samples when only two categories, euploid or aneuploid/mosaic, were grouped to calculate the concordance.</p></sec><sec><title>Conclusions</title><p>Compared with BF, SCM has superior diagnostic performance, and it is non-invasive for embryos.</p></sec><sec><title>Clinical Trial Registration</title><p>[<ext-link ext-link-type="uri" xlink:href="http://www.chictr.org.cn" xmlns:xlink="http://www.w3.org/1999/xlink">http://www.chictr.org.cn</ext-link>], identifier [ChiCTR-BPD-17014087].</p></sec>