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

• 資料種別: 博士論文
• タイトル: Isolation and characterization of fetal nucleated red blood cells from maternal blood as a target for single cell sequencing-based non-invasive genetic testing
• 著者・編者: 伊東, 紀子
• 著者標目: 伊東, 紀子
• 出版事項: Japan Society for Reproductive Medicine
• 出版年月日等: 2021-09-01
• 出版年（W3CDTF）: 2021-09-01
• 並列タイトル等: 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
• タイトル（掲載誌）: Reproductive Medicine and Biology
• 巻号年月日等（掲載誌）: 20 3
• 掲載巻: 20
• 掲載号: 3
• 掲載ページ: 352-360
• ISSN（掲載誌）: ISSN : 1447-0578
• 授与機関名: Nagasaki University (長崎大学)
• 授与年月日: 2021-09-01
• 報告番号: 甲医歯薬第1366号
• 学位: 博士(薬学)
• 本文の言語コード: eng
• 件名標目: fetal nucleated red blood cell (fNRBC); fluorescence-activated; cell sorting (FACS), non-invasive; prenatal testing (NIPT), single cell, whole genome amplification (WGA)
• 対象利用者: 一般
• 一般注記: 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)
• 一次資料へのリンクURL: https://nagasaki-u.repo.nii.ac.jp/?action=repository_action_common_download&item_id=27087&item_no=1&attribute_id=70&file_no=1 （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
• オンライン閲覧公開範囲: インターネット公開
• 著作権情報: © 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.
• 関連情報: http://hdl.handle.net/10069/00040953
• 関連情報（DOI）: 10.1002/rmb2.12392
• 連携機関・データベース: 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
• 提供元機関・データベース: 長崎大学 : 長崎大学学術研究成果リポジトリ