RT-qPCR analyses on the osteogenic differentiation from human iPS cells: an investigation of reference genes.

• Material Type: 博士論文
• Title: RT-qPCR analyses on the osteogenic differentiation from human iPS cells: an investigation of reference genes.
• Author/Editor: Okamura, Kensuke; Inagaki, Yusuke; Matsui, Takeshi K.; Matsubayashi, Masaya; Komeda, Tomoya; Ogawa, Munehiro; Mori, Eiichiro; Tanaka, Yasuhito
• Author Heading: Okamura, Kensuke; Inagaki, Yusuke; Matsui, Takeshi K.; Matsubayashi, Masaya; Komeda, Tomoya; Ogawa, Munehiro; Mori, Eiichiro; Tanaka, Yasuhito
• Publication, Distribution, etc.: Nature Publishing Group
• Publication Date: 2020-07-16
• Publication Date (W3CDTF): 2020-07-16
• Alternative Title: ヒトiPS細胞の骨分化誘導における逆転写-定量的リアルタイムPCR (RT-qPCR) で用いる参照遺伝子についての検討
• Periodical title: Scientific reports
• No. or year of volume/issue: 10 1
• Volume: 10
• Issue: 1
• Pages: Article No.11748-
• ISSN (Periodical Title): ISSN : 2045-2322
• Degree grantor/type: 奈良県立医科大学
• Date Granted: 2021-03-15
• Dissertation Number: 甲第770号
• Degree Type: 博士（医学）
• Text Language Code: eng
• Subject Heading: Molecular biology; Stem cells
• Target Audience: 一般
• Note (General): type:Thesis; Reverse transcription quantitative PCR (RT-qPCR) is used to quantify gene expression and require standardization with reference genes. We sought to identify the reference genes best suited for experiments that induce osteogenic differentiation from human induced pluripotent stem cells. They were cultured in an undifferentiated maintenance medium and after confluence, further cultured in an osteogenic differentiation medium for 28 days. RT-qPCR was performed on undifferentiation markers, osteoblast and osteocyte differentiation markers, and reference gene candidates. The expression stability of each reference gene candidate was ranked using four algorithms. General rankings identified TATA box binding protein in the first place, followed by transferrin receptor, ribosomal protein large P0, and finally, beta-2-microglobulin, which was revealed as the least stable. Interestingly, universally used GAPDH and ACTB were found to be unsuitable. Our findings strongly suggest a need to evaluate the expression stability of reference gene candidates for each experiment.; 博士（医学）・甲第770号・令和3年3月15日; © The Author(s) 2020 : Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.; identifier:Scientific reports Vol.10 No.1 Article No.11748 (2020 Jul); identifier:20452322; identifier:http://ginmu.naramed-u.ac.jp/dspace/handle/10564/3894; identifier:Scientific reports, 10(1): Article No.11748
• Source: fulltext; http://ginmu.naramed-u.ac.jp/dspace/bitstream/10564/3894/1/01%e7%94%b2770%e6%9c%ac%e6%96%87%e3%81%ae%e8%a6%81%e6%97%a8.pdf; http://ginmu.naramed-u.ac.jp/dspace/bitstream/10564/3894/2/02%e7%94%b2770%e5%af%a9%e6%9f%bb%e8%a6%81%e6%97%a8.pdf; http://ginmu.naramed-u.ac.jp/dspace/bitstream/10564/3894/3/03%e7%94%b2770%e6%9c%ac%e6%96%87.pdf
• Access Restrictions: インターネット公開
• Related Material (DOI): 10.1038/s41598-020-68752-2
• 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): 奈良県立医科大学 : 奈良県立医科大学機関リポジトリ GINMU