Establishment of FACS-mQ as a tool for detecting stem cells or cancer stem cells

• Project/Area Number: 23590670
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Laboratory medicine
• Research Institution: Osaka University
• Principal Investigator: TAKANO Toru 大阪大学, 医学(系)研究科(研究院), 講師 (00263236)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2011: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: FACS-mQ / flow cytometry / 幹細胞 / 癌幹細胞 / RNA / 遺伝子発現プロフィール / FACS-ｍQ / Flow Cytometry

Research Abstract

The basic protocols of FACS-mQ(mRNA quantification after FACS) were established. In FACS-mQ, intracellular RNAs are preserved without degradation after immunolabeling with a florophore, then we can analyze gene expression profiles of sorted cells after FACS. We established the following three protocols: 1)preparation of single cells from blood or tissue samples without RNA degradation, 2)high sensitive immunolabeling of the targeted cells, and 3) recovery and analysis of a samll amount of RNAs from sorted cells.

Research Products

• [Journal Article] An improved protocol for mRNA quantification after fluorescence activated cell sorting with an increased signal to noise ratio in flow cytometry (2014)
  • Author(s): Date, A., Maeda, T., Watanabe, M., Hidaka, Y., Iwatani, Y., Takano, T
  • Journal Title: Mol Biotechnol Volume: (in press) Issue: 7 Pages: 591-598
  • DOI: 10.1007/s12033-014-9733-5
  • Peer Reviewed
• [Journal Article] Fetal cell carcinogenesis of the thyroid : A modified theory based on recent evidence (2014)
  • Author(s): Takano, T
  • Journal Title: Endocr J Volume: 61(DN/JST.JSTAGE/endocrj/EJ13-0517 [pii]) Pages: 311-320
  • Peer Reviewed
• [Journal Article] Fetal cell carcinogenesis of the thyroid: A modified theory based on recent evidence [My Opinion] (2014)
  • Author(s): Toru Takano
  • Journal Title: Endocrine Journal Volume: 61 Issue: 4 Pages: 311-320
  • DOI: 10.1507/endocrj.EJ13-0517
  • NAID: 130004443957
  • ISSN: 0918-8959, 1348-4540
  • Peer Reviewed
• [Journal Article] Genes that characterize T3-predominant Graves' thyroid tissues (2013)
  • Author(s): Matsumoto, C., Ito, M., Yamada, H., Yamakawa, N., Yoshida, H., Date, A., Watanabe, M., Hidaka, Y., Iwatani, Y., Miyauchi, A., Takano, T
  • Journal Title: Eur J Endocrinol Volume: 168(2) Issue: 2 Pages: 137-44
  • DOI: 10.1530/eje-12-0507
  • Peer Reviewed
• [Journal Article] Preparation of thyroid follicular cells for mRNA quantifi cation after fl uorescence-activated cell sorting (2013)
  • Author(s): Matsumoto, C., Ito, M., Yamada, H., Yoshida, H., Watanabe, M., Hidaka, Y., Iwatani, Y., Miyauchi, A., Takano, T
  • Journal Title: Scandinavian Journal of Clinical & Laboratory Investigation Volume: 73 Issue: 3 Pages: 245-252
  • DOI: 10.3109/00365513.2013.769624
  • Peer Reviewed
• [Journal Article] Prolonged hybridization with a cRNA probe improves the signal to noise ratio for in-tube in situ hybridization for quantification of mRNA after fluorescence-activated cell sorting (2012)
  • Author(s): Yamada, H., Yamakawa, N., Watanabe, M., Hidaka, Y., Iwatani, Y., Takano, T.
  • Journal Title: Biotech Histochem Volume: (in press) Issue: 5 Pages: 366-71
  • DOI: 10.3109/10520295.2012.672650
  • Peer Reviewed
• [Journal Article] Measurement of TFF3 mRNA in aspirates from thyroid nodules using mesh filtration : The first clinical trial in 130 cases (2012)
  • Author(s): Yamada, H., Takano, T., Kihara, M., Hirokaw,a M., Yoshida, H., Watanabe, M., Iwatani, Y., Hidaka, Y., .Miyauchi, A
  • Journal Title: Endocr J Volume: 59(JST.JSTAGE/endocrj/EJ12-0077 [pii]) Pages: 621-630
  • Peer Reviewed
• [Journal Article] Measurement of TFF3 mRNA in aspirates from thyroid nodules using mesh filtration: The first clinical trial in 130 cases. (2012)
  • Author(s): .Yamada, H., Takano, T., Kihara, M., Hirokaw,a M., Yoshida, H., Watanabe, M., Iwatani, Y., Hidaka, Y., .Miyauchi, A.
  • Journal Title: Endocr J Volume: 59 Pages: 621-630
  • Peer Reviewed
• [Journal Article] mRNA quantification after fluorescence activated cell sorting using locked nucleic acid probes (2011)
  • Author(s): Maruo, R., Yamada, H., Watanabe, M., Hidaka, Y., Iwatani, Y. Takano, T.
  • Journal Title: Mol Biotechnol Volume: 49(1) Issue: 1 Pages: 42-47
  • DOI: 10.1007/s12033-011-9375-9
  • Peer Reviewed
• [Presentation] Preoperative molecular diagnosis of thyroid nodule。How does it help? (2014)
  • Author(s): Toru Takano
  • Organizer: The ３ｒｄ Asian Masterclass of Thyroid Cancer
  • Place of Presentation: Taipei, Taiwan
  • Invited
• [Presentation] Preoperative molecular diagnosis of thyroid nodule (2013)
  • Author(s): Takano, T
  • Organizer: The 2nd Asian Masterclass of Thyroid Cancer
  • Place of Presentation: Soul, South Korea
  • Year and Date: 2013-05-25
  • Invited
• [Presentation] 甲状腺濾胞癌の分子診断 (2012)
  • Author(s): 高野徹
  • Organizer: 第53回日本臨床細胞学会総会
  • Place of Presentation: 千葉
  • Year and Date: 2012-06-02
• [Presentation] 新規幹細胞解析法、FACS-mQによる甲状腺幹細胞・癌幹細胞の同定・解析 (2011)
  • Author(s): 髙野徹
  • Organizer: 第54回日本甲状腺学会
  • Place of Presentation: 大阪
  • Year and Date: 2011-11-22
• [Presentation] 新規臨床検査法、FACS-mQの開発 (2011)
  • Author(s): 髙野徹, 日高洋, 岩谷良則
  • Organizer: 第58回日本臨床検査医学会学術集会
  • Place of Presentation: 岡山
  • Year and Date: 2011-11-18
• [Presentation] 甲状腺腫瘍に対する穿刺吸引核酸診断法の開発 (2011)
  • Author(s): 髙野徹
  • Organizer: 第54回日本臨床検査医学会近畿支部総会
  • Place of Presentation: 滋賀
  • Year and Date: 2011-10-29
• [Presentation] 新規臨床検査法、FACS-ｍQの開発
  • Author(s): 高野 徹、日高 洋、岩谷 良則
  • Organizer: 第58回日本臨床検査医学会学術集会(招待講演)
  • Place of Presentation: 岡山
• [Presentation] 新規幹細胞解析法、FACS-mQによる甲状腺幹細胞・癌幹細胞の同定・解析
  • Author(s): 高野 徹
  • Organizer: 第８４回日本甲状腺学会(招待講演)
  • Place of Presentation: 大阪
• [Remarks]
  • URL: http://www.med.osaka-u.ac.jp/pub/labo/www/CRT/CRT%20Home.html
• [Remarks] 大阪大学大学院医学系研究科臨床検査診断学腫瘍研究チーム
  • URL: http://www.med.osaka-u.ac.jp/pub/labo/www/CRT/CRT%20Home.html
• [Remarks] 大阪大学大学院医学系研究科臨床検査診断学腫瘍研究チーム
  • URL: http://www.med.osaka-u.ac.jp/pub/labo/www/CRT/CRT%20Home.html
• [Remarks]
  • URL: http://www.med.osaka-u.ac.jp/pub/labo/www/CRT/CRT%20Home.html