دورية أكاديمية
أعرض في EDS

A workflow for simultaneous DNA copy number and methylome analysis of inner cell mass and trophectoderm cells from human blastocysts.

التفاصيل البيبلوغرافية
العنوان: A workflow for simultaneous DNA copy number and methylome analysis of inner cell mass and trophectoderm cells from human blastocysts.
المؤلفون: Olcha M; Department of Obstetrics and Gynecology, Montefiore Medical Center/Albert Einstein College of Medicine, Bronx, New York. Electronic address: olchame@gmail.com., Dong X; Department of Genetics, Albert Einstein College of Medicine, Bronx, New York., Feil H; Department of Obstetrics and Gynecology, Montefiore Medical Center/Albert Einstein College of Medicine, Bronx, New York., Hao X; Department of Genetics, Albert Einstein College of Medicine, Bronx, New York., Lee M; Department of Genetics, Albert Einstein College of Medicine, Bronx, New York., Jindal S; Department of Obstetrics and Gynecology, Montefiore Medical Center/Albert Einstein College of Medicine, Bronx, New York., Buyuk E; Department of Obstetrics and Gynecology, Montefiore Medical Center/Albert Einstein College of Medicine, Bronx, New York; Reproductive Medicine Associates of New York, Division of Reproductive Endocrinology and Infertility, Department of Obstetrics, Gynecology, and Reproductive Science, Icahn School of Medicine at Mount Sinai, New York, New York., Vijg J; Department of Genetics, Albert Einstein College of Medicine, Bronx, New York; Center for Single-Cell Omics in Aging and Disease, School of Public Health, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.
المصدر: Fertility and sterility [Fertil Steril] 2021 Jun; Vol. 115 (6), pp. 1533-1540. Date of Electronic Publication: 2021 Feb 12.
نوع المنشور: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't
اللغة: English
بيانات الدورية: Publisher: Elsevier for the American Society for Reproductive Medicine Country of Publication: United States NLM ID: 0372772 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1556-5653 (Electronic) Linking ISSN: 00150282 NLM ISO Abbreviation: Fertil Steril Subsets: MEDLINE
أسماء مطبوعة: Publication: New York. NY : Elsevier for the American Society for Reproductive Medicine
Original Publication: New York, Hoeber.
مواضيع طبية MeSH: DNA Copy Number Variations* , DNA Methylation* , Epigenesis, Genetic* , Epigenome* , Epigenomics* , Gene Dosage* , Single-Cell Analysis*, Blastocyst Inner Cell Mass/*pathology , Trophoblasts/*pathology, Aneuploidy ; Blastocyst Inner Cell Mass/metabolism ; Cell Separation ; CpG Islands ; Female ; Gene Expression Regulation, Developmental ; Humans ; Trophoblasts/metabolism ; Whole Genome Sequencing ; Workflow
مستخلص: Objective: To establish a workflow for isolating single trophectoderm (TE) and inner cell mass (ICM) cells and to simultaneously evaluate these cells for copy number variation (CNV) as well as methylome development.
Design: Experimental.
Setting: Academic medical center.
Patient(s): Donated genetically abnormal blastocysts.
Intervention(s): Single cells were isolated, followed by bisulfite conversion and sequencing to identify CNV and methylome profiles.
Main Outcome Measure(s): CNV and methylation profiling.
Result(s): Two embryos were dissociated, isolating 46 single cells, with 17 ICM and 12 TE cells selected for further downstream analysis. Chromosome ploidies and embryo sex were concordant with the results from conventional aneuploidy testing. In 3 of the 29 cells, additional aneuploidies were discovered, indicating possible mosaicism undetected by routine preimplantation genetic testing for aneuploidy. CpG methylation frequency was higher in ICM cells compared with TE cells (44.3% vs. 32.4%), respectively, while non-CpG methylation frequency was similar among both cell types. CpG methylation levels accurately distinguished ICM from TE cells epigenetically.
Conclusion(s): We describe an effective workflow for isolating and sequencing single ICM and TE cells from human blastocysts. The use of methylation profiling can help distinguish these two cell populations better then morphologic identification alone. TE cells had significantly lower levels of DNA methylation, which may be explained in part by the fact that these cells have begun the process of differentiation and are transcriptionally more active than ICM. This approach may be used to explore the genetic complexities within human embryos, specifically among the two primary cell types seen at this stage of development.
(Copyright © 2020 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)
التعليقات: Comment in: Fertil Steril. 2021 Jun;115(6):1441-1442. (PMID: 34053516)
فهرسة مساهمة: Keywords: PGT-A; methylation; single cell sequencing
تواريخ الأحداث: Date Created: 20210216 Date Completed: 20210823 Latest Revision: 20210823
رمز التحديث: 20240628
DOI: 10.1016/j.fertnstert.2020.11.007
PMID: 33589136
قاعدة البيانات: MEDLINE
الوصف
تدمد:1556-5653
DOI:10.1016/j.fertnstert.2020.11.007