並列タイトル等ヒト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
一般注記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
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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
DOIinfo:doi/10.1038/s41598-020-68752-2
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