Isolation and characterization of fetal nucleated red blood cells from maternal blood as a target for single cell sequencing-based non-invasive genetic testing

• Material Type: 博士論文
• Title: Isolation and characterization of fetal nucleated red blood cells from maternal blood as a target for single cell sequencing-based non-invasive genetic testing
• Author/Editor: 伊東, 紀子
• Author Heading: 伊東, 紀子
• Publication, Distribution, etc.: Japan Society for Reproductive Medicine
• Publication Date: 2021-09-01
• Publication Date (W3CDTF): 2021-09-01
• Alternative Title: 母体血中の胎児有核赤血球細胞を標的としたシングルセルシーケンスによる出生前遺伝学的検査法の開発
• Periodical title: Reproductive Medicine and Biology
• No. or year of volume/issue: 20 3
• Volume: 20
• Issue: 3
• Pages: 352-360
• ISSN (Periodical Title): ISSN : 1447-0578
• Degree grantor/type: Nagasaki University (長崎大学)
• Date Granted: 2021-09-01
• Dissertation Number: 甲医歯薬第1366号
• Degree Type: 博士(薬学)
• Text Language Code: eng
• Subject Heading: fetal nucleated red blood cell (fNRBC); fluorescence-activated; cell sorting (FACS), non-invasive; prenatal testing (NIPT), single cell, whole genome amplification (WGA)
• Target Audience: 一般
• Note (General): Purpose: Although non-invasive prenatal testing (NIPT) based on cell-free DNA (cfDNA) in maternal plasma has been prevailing worldwide, low levels of fetal DNA fraction may lead to false-negative results. Since fetal cells in maternal blood provide a pure source of fetal genomic DNA, we aimed to establish a workflow to isolate and sequence fetal nucleated red blood cells (fNRBCs) individually as a target for NIPT.Methods: Using male-bearing pregnancy cases, we isolated fNRBCs individually from maternal blood by FACS, and obtained their genomic sequence data through PCR screening with a Y-chromosome marker and whole-genome amplification (WGA)-based whole-genome sequencing.Results: The PCR and WGA efficiencies of fNRBC candidates were consistently lower than those of control cells. Sequencing data analyses revealed that although the majority of the fNRBC candidates were confirmed to be of fetal origin, many of the WGA-based genomic libraries from fNRBCs were considered to have been amplified from a portion of genomic DNA.Conclusions: We established a workflow to isolate and sequence fNRBCs individually. However, our results demonstrated that, to make cell-based NIPT targeting fNRBCs feasible, cell isolation procedures need to be further refined such that the nuclei of fNRBCs are kept intact.; 長崎大学学位論文 学位記番号:博(医歯薬)甲第1366号 学位授与年月日:令和3年9月1日; Author: Noriko Ito, Kazuhiro Tsukamoto, Kosuke Taniguchi, Ken Takahashi, Aikou Okamoto, Hiroaki Aoki, Yuka Otera-Takahashi, Michihiro Kitagawa, Hiroko Ogata-Kawata, Hideaki Morita, Kenichiro Hata, Kazuhiko Nakabayashi; Citation: Reproductive Medicine and Biology, 20(3), pp.352-360; 2021; identifier:Nagasaki University (長崎大学), 博士(薬学) (2021-09-01)
• Source: fulltext; https://nagasaki-u.repo.nii.ac.jp/?action=repository_action_common_download&item_id=27087&item_no=1&attribute_id=70&file_no=1
• Access Restrictions: インターネット公開
• Rights (production): © 2021 The Authors. Reproductive Medicine and Biology published by John Wiley & Sons Australia, Ltd on behalf of Japan Society for Reproductive Medicine. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
• Related Material: http://hdl.handle.net/10069/00040953
• Related Material (DOI): 10.1002/rmb2.12392
• Data Provider (Database): 国立情報学研究所 : 学術機関リポジトリデータベース（IRDB）（機関リポジトリ）; https://irdb.nii.ac.jp
• Original Data Provider (Database): 長崎大学 : 長崎大学学術研究成果リポジトリ