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- W2751957570 abstract "Next generation sequencing (NGS) has the ability to detect ratios of normal:abnormal cells in blastocyst biopsies. With the current biopsy methodology clumps of 4-6 cells are analyzed together and a 50:50 mixture of monosomic:trisomic cells would be reported as a normal embryo. However, scoring a single clump of cells cannot reliably identify the true degree of mosaicism in trophectoderm samples [1]. The purpose of this study was to dissect single-cells from human blastocyst biopsies and to prove that it is possible to successfully carry out NGS analysis. Research prospective study. Two human blastocysts donated for research (IRB HIC#2000020219) were enzymatically dissected into single cells and each cell was individually isolated and snap frozen in LN2. A total of sixty (n=60) single cells were collected for this study, and 33 cells were randomly chosen for NGS library preparation (5 from one embryo and 28 from the other). A single collection of 4-6 cells from the same embryos were considered the control group (standard biopsy). DNA from single cells was amplified by SurePlex, sequenced with VeriSeq PGS assay (Illumina) on MiSeq (Illumina) and analyzed with BlueFuse Multi analysis (Illumina). Most cells from both embryos were successfully disaggregated and appeared alive. Successful amplification was obtained from 23 (70%) out of 33 single cells selected for the study. The average reads obtained per cell was 357,180. Interestingly, good concordance was shown between the control and the single cells collected from the same embryo (83% concordance rate) and between cells (85%). This study proves the first successful disaggregation of trophectoderm from human blastocysts into single cells and subsequent NGS analysis. This technique will be helpful in: a) establishing the true magnitude of embryo mosaicism; b) providing a more robust tool (analysis of single cells as opposed to clumps) for PGS analysis in the context of IVF treatments; c) clarifying the extent of concordance of genetic information within the embryo; and d) determining the type of aneuploidy/mosaicism among cells." @default.
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- W2751957570 date "2017-09-01" @default.
- W2751957570 modified "2023-10-14" @default.
- W2751957570 title "A novel method for single cell dissection and sequencing of human blastocysts" @default.
- W2751957570 doi "https://doi.org/10.1016/j.fertnstert.2017.07.820" @default.
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