CReATe Fertility Center, Toronto, Canada.
Department of Obstetrics and Gynecology, Faculty of Medicine, University of Toronto, Toronto, ON, Canada.
PLoS One. 2018 May 10;13(5):e0197262. doi: 10.1371/journal.pone.0197262. eCollection 2018.
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).
47 embryos from 35 patients undergoing IVF.
DNA analysis of combined BCCM plus BF in comparison with trophectoderm (TE) biopsy and/or whole blastocyst (WB)using next generation sequencing (NGS).
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).
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.
评估从囊胚培养条件培养基（BCCM）中分离的胚胎 DNA 与囊胚腔液（BF）结合是否可用于囊胚期非侵入性植入前遗传学检测染色体非整倍体（非侵入性植入前遗传学筛查，NIPGS）。
35 名接受体外受精的患者的 47 个胚胎。
与滋养外胚层（TE）活检和/或整个囊胚（WB）比较，对 BCCM 加 BF 进行 DNA 分析，使用下一代测序（NGS）。
在 47/47 个 NIPGS 样本（28 个冷冻解冻和 19 个新鲜培养样本）中成功扩增了胚胎 DNA，范围为 6.3 至 44.0 ng/μl。对于冷冻解冻的胚胎，每个样本的全染色体拷贝数的一致性率在 NIPGS 与 TE 活检之间，NIPGS 与 WB 之间以及取自同一胚胎的 TE 与 WB 之间分别为 87.5%；96.4%和 91.7%（P>0.05），每个单染色体的一致性率分别为 99.3%，99.7%和 99.7%（P>0.05）。在新鲜病例（第 4 天至第 5/6 天培养）中，取自同一胚胎的 NIPGS 与 TE 样本之间的全染色体拷贝数的一致性率为 100%，每个单染色体的一致性率为 98.2%（P>0.05）。
BCCM 和 BF 的组合包含足够的胚胎 DNA 用于全基因组扩增和准确的非整倍体筛查。我们的研究结果表明，使用 BCCM 与 BF 联合进行非整倍体筛查可能成为人类 IVF 的一种新的 NIPGS 方法。
