Utility of separation technique of live cancer stem cells with using an energy transfer fluorescence probes

• Project/Area Number: 21590622
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Laboratory medicine
• Research Institution: Hamamatsu University School of Medicine
• Principal Investigator: TAKESHITA Akihiro 浜松医科大学, 医学部, 准教授 (00242769)
• Co-Investigator(Kenkyū-buntansha): OHNISHI Kazunori 浜松医科大学, 医学部附属病院, 教授 (80252170) ; 竹下 香 (新庄 香) 浜松医科大学, 保健管理センター, 特任助教 (30397393)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2011: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2010: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2009: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
• Keywords: エネルギー移行 / cell sorter / mRNA / 白血病 / 幹細胞 / FRET法 / KRAS / 癌細胞 / 白血病細胞 / bcr-abl / Kras / 肝細胞

Research Abstract

QUAL/FRET method is effective to determine the cells in which have specific mRNA. In this study, we examined the efficacy on insertion of probes using fluorescein labeled dextran firstly. Relatively high molecular ones did not enter into nucleus and distributed in cytoplasm. On the other hand, the efficacy of the insertion was studied in cell lines and peripheral blood cells. The efficacy was not sufficient in adherent cells and lymphocytes. It should be improved by changing molecular size of probe and conjugated fluorescence.

Research Products

• [Journal Article] Japan Adult Leukemia Study Group. Impact of additional chromosomal abnormalities in patients with acute promyelocytic leukemia : 10-year results of the Japan Adult Leukemia Study Group APL97study (2011)
  • Author(s): Ono T, Takeshita A, Iwanaga M, Asou N, Naoe T, Ohno R
  • Journal Title: Haematologica Volume: 96 Pages: 174-176
• [Journal Article] for the Japan Adult Leukemia Study Group. Phase I trial of gemtuzumab ozogamicin in intensive combination chemotherapy for relapsed or refractory adult acute myeloid leukemia(AML): Japan Adult Leukemia Study Group(JALSG)-AML206 study (2011)
  • Author(s): Usui N, Takeshita A, Nakaseko C, Dobashi N, Fujita H, Kiyoi H, Kobayashi Y, Sakura T, Yahagi Y, Shigeno K, Ohwada C, Miyazaki Y, Ohtake S, Miyawaki S, Naoe T, Ohnishi K
  • Journal Title: Cancer Sci Volume: 102 Pages: 1358-1365
• [Journal Article] Phase I trial of gemtuzumab ozogamicin in intensive combination chemotherapy for relapsed or refractory adult acute myeloid leukemia (AML) : Japan Adult Leukemia study Group (JALSG)-AML206 study (2011)
  • Author(s): Usui N, Ohnishi K, et al
  • Journal Title: Cancer Science Volume: 102(7) Issue: 7 Pages: 1358-65
  • DOI: 10.1111/j.1349-7006.2011.01957.x
  • NAID: 10029295449
  • Peer Reviewed
• [Journal Article] Impact of additional chromosomal abnormalities in patients with acute promyelocytic leukemia : 10-year results of the Japan Adult Leukemia Study Group APL97 study (2011)
  • Author(s): Ono T, Takeshita A, et al
  • Journal Title: Haematologica Volume: 96(1) Issue: 1 Pages: 174-6
  • DOI: 10.3324/haematol.2010.030205
  • Peer Reviewed
• [Journal Article] MEDICAL TECHNOLOGISTS AT UNIVERSITY HOSPITAL BLOOD TRANSFUSION DEPARTMENTS PLAY AN IMPORTANT ROLE IN EDUCATION IN TRANSFUSION MEDICINE: THE 2009 TRANSFUSION CONFERENCE OF JAPANESE UNIVERSITY HOSPITALS. SURVEILLANCE REPORT ON MEDICAL EDUCATION (1) (2011)
  • Author(s): Fujihara H, Takeshita A, et al
  • Journal Title: Japanese Journal of Transfusion and Cell Therapy Volume: 57 Issue: 6 Pages: 470-477
  • DOI: 10.3925/jjtc.57.470
  • NAID: 130004542649
  • ISSN: 1881-3011, 1883-0625
  • Peer Reviewed
• [Journal Article] Impact of additional chromosomal abnormalities in patients with acute promyelocytic leukemia : 10-year results of the Japan Adult Leukemia Study Group APL97 study. (2011)
  • Author(s): Ono T, Takeshita A, et al
  • Journal Title: Haematologica Volume: 96(1) Pages: 174-176
  • Peer Reviewed
• [Journal Article] Collaborative study of irregular erythrocyte antibodies in Japan : results from the Japanese study group of allo-immunity and antigen diversity in Asian populations (2010)
  • Author(s): Takeshita A, Watanabe H, Fijihara H, Oshida M, Yurugi K, Tomoda Y, Uchikawa M, Kino S, Ohto H
  • Journal Title: Transfus Apher Sci Volume: 43 Pages: 3-8
• [Journal Article] Collaborative study of irregular erythrocyte antibodies in Japan : results from the Japanese study group of allo-immunity and antigen diversity in Asian populations. (2010)
  • Author(s): Takeshita A, et al
  • Journal Title: Transfus Apheresis Sciences Volume: 43(1) Pages: 3-8
  • Peer Reviewed
• [Journal Article] CMC-544(inotuzumab ozogamicin) shows less effect on multidrug resistant cells : analyses in cell lines and cells from patients with B-cell chronic lymphocytic leukaemia and lymphoma (2009)
  • Author(s): Takeshita A, Shinjo K, Yamakage N, Ono T, Hirano I, Matsui H, Shigeno K, Nakamura S, Tobita T, Maekawa M, Ohnishi K, Sugimoto Y, Kiyoi H, Naoe T, Ohno R
  • Journal Title: Br J Haematol Volume: 146(1) Pages: 34-43
• [Journal Article] Effective Blood Utilization via System for Massive Blood Transfusion, including Cardiovascular Operation in Local Areas (2009)
  • Author(s): Takeshita A, Asai T, Murakami M, Fujihara H, Ishizuka T, Nakai S, Yamada C, Suzumura T, Uchiyama Y, Maekawa M, Shigeno K, Washiyama N, Yamashita K, Unno N, Shinjo K
  • Journal Title: Jpn J Transfusion Cell Therapy Volume: 55(1) Pages: 63-7
  • NAID: 10025628900
• [Journal Article] CMC-544(inotuzumab ozogamicin), an anti-CD22immuno-conjugate of calicheamicin, alters the levels of target molecules of malignant B-cells (2009)
  • Author(s): Takeshita A, Yamakage N, Shinjo K, Ono T, Hirano I, Nakamura S, Shigeno K, Tobita T, Maekawa M, Kiyoi H, Naoe T, Ohnishi K, Sugimoto Y, Ohno R
  • Journal Title: Leukemia Volume: 23(7) Pages: 1329-36
• [Journal Article] Phase I/II study of humanized anti-CD33antibody conjugated with calicheamicin, gemtuzumab ozogamicin, in relapsed or refractory acute myeloid leukemia : final results of Japanese multicenter cooperative study (2009)
  • Author(s): Kobayashi Y, Tobinai K, Takeshita A, Naito K, Asai O, Dobashi N, Furusawa S, Saito K, Mitani K, Morishima Y, Ogura M, Yoshiba F, Hotta T, Bessho M, Matsuda S, Takeuchi J, Miyawaki S, Naoe T, Usui N, Ohno R
  • Journal Title: Int J Hematol Volume: 89(4) Pages: 460-9
  • NAID: 10025161243
• [Journal Article] CMC-544 (inotuzumab ozogamicin) shows less effect on multidrug resistant cells : analyses in cell lines and cells from patients with B-cell chronic lymphocytic leukaemia and lymphoma (2009)
  • Author(s): Takeshita A, et al.
  • Journal Title: Br J Haematol 146 Pages: 34-43
  • Peer Reviewed
• [Journal Article] CMC-544 (inotuzumab ozogamicin), an anti-CD22 immuno-conjugate of calicheamicin, alters the levels of target molecules of malignant B-cells (2009)
  • Author(s): Takeshita A, et al.
  • Journal Title: Leukemia 23 Pages: 1329-1336
  • Peer Reviewed
• [Journal Article] Phase I/II study of humanized anti-CD33 antibody conjugated with calicheamicin, gemtuzumab ozogamicin, in relapsed or refractory acute myeloid leukemia : final results of Japanese multicenter cooperative study (2009)
  • Author(s): Kobayashi Y, et al.
  • Journal Title: Int J Hematol 89 Pages: 460-469
  • Peer Reviewed
• [Journal Article] Effective Blood Utilization via System for Massive Blood Transfusion, including Cardiovascular Operation in Local Areas (2009)
  • Author(s): Takeshita A, et al.
  • Journal Title: Jpn J Transfusion Cell Therapy 55 Pages: 63-7
  • NAID: 10025628900
  • Peer Reviewed
• [Journal Article] Education in transfusion medicine for medical students and doctors
  • Author(s): Takeshita A, Yoneyama Y, Fujihara H, Nollet KE, Ohto H
  • Journal Title: Vox Sanguinis Volume: (in press)
• [Presentation] Efficacy of gemtuzumab ozogamicin(GO) on relapsed/refractory acute promyelocytic leukemia(APL) (2011)
  • Author(s): Takeshita A, Ono T, Kojima Y, Kyo T, Asou N, Suzushima H, Yagasaki H, Maeda T, Okada M, Nakaseko C, Kanbayashi H, Yamaguchi M, Kurosawa M, Tsuboi K, Yjiri T, Nanya Y, UikeN, Akiyama N, Fukuda T, Karimata K, Okumura H, Eto T, Izumiyama K, Watanabe A, Aotsuka N, Ito K, Ozaki K, Naoe T
  • Organizer: 53rd Annual Meeting of American Society of Hematology San Diego
  • Place of Presentation: CA, USA
  • Year and Date: 2011-12-10
• [Presentation] Differences in allo-immunity to erythrocytes between Korea and Japan. Cooperative international study of alloimmunity to antigen diversity in Asian population (2011)
  • Author(s): Fujihara H, Takeshita A, Kim DW, Han KS, Kwon SY, Kwon SW, Suh JS, Watanabe H, Uchikawa M, Kino S, Nollet KE, Ohto H
  • Organizer: 53rd Annual Meeting of American Society of Hematology San Diego
  • Place of Presentation: CA, USA
  • Year and Date: 2011-12-10
• [Presentation] Clinical features and prognostic impact of CD56expression in acute promyelocytic leukemia : Long term follow up data from the Japan Adult Leukemia Study Group(JALSG) APL97 (2011)
  • Author(s): Ono T, Takeshita A, Kishimoto Y, Kiyoi H, Okada M, Yamauchi T, Tsuzuki M, Horikawa K, Matsuda M, Shinagawa K, Monma F, Ohtake S, Nakaseko C, Takahashi M, Yagasaki F, Kimura Y, Fujita H, Iwanaga M, Asou N, Ohnishi K, Naoe T
  • Organizer: 53rd Annual Meeting of American Society of Hematology San Diego
  • Place of Presentation: CA, USA
  • Year and Date: 2011-12-10
• [Presentation] Role of hematopoietic stem cell transplantation as salvage treatment of acute promyelocytic leukemia initially treated with all-trans retinoic acid : a retrospective analysis of the Japan Adult Leukemia Study Group(JALSG) APL97Study (2011)
  • Author(s): Fujita H, Asou N, Iwanaga M, Hyo R, Nomura S, Kiyoi H, Okada M, Tsuzuki M, Matsuda M, Yamauchi T, Ohtake S, Izumi T, Nakaseko C, Mitani K, Shinagawa K, Takeshita A, Miyazaki Y, Ohnishi K, Miyawaki S, Naoe T
  • Organizer: 53rd Annual Meeting of American Society of Hematology San Diego
  • Place of Presentation: CA, USA
  • Year and Date: 2011-12-10
• [Presentation] The usefulness and demerit of the expected elevation value of serum albumin before replacement therapy (2011)
  • Author(s): Fujihara H, Yamada C, Watanabe H, Shibata H, Funai Y, Suzumura T, Kaneko M, Uchiyama S, Takeshita A
  • Organizer: 16th Congress of the European Hematology Association
  • Place of Presentation: London, UK
  • Year and Date: 2011-06-09
• [Presentation] Allo-Immunity to Erythrocytes in Relation to Blood Transfusion and Pregnancy ; Report from the Asian Cooperative Group (2011)
  • Author(s): Takeshita A
  • Organizer: 30^<th> Annual Meeting of Korean Society of Blood Transfusion
  • Place of Presentation: Busan, Korea
  • Year and Date: 2011-06-09
• [Presentation] Our role in promoting a better understanding of the efficacy of cryoprecipitate in acquired hypofibrinoginemia induced by perioperative massive bleeding (2011)
  • Author(s): Yamada C, Fujihara H, Watanabe H, Takeshita A
  • Organizer: International Society of Blood Transfusion
  • Place of Presentation: Lisbon, Portugal
  • Year and Date: 2011-06-06
• [Presentation] Allo-Immunity to Erythrocytes in Relation to Blood Transfusion and Pregnancy ; Report from the Asian Cooperative Group (2011)
  • Author(s): Takeshita A, et al
  • Organizer: 30^<th> Annual Meeting of Korean Society of Blood Transfusion
  • Place of Presentation: Busa, Korea
• [Presentation] Efficacy of gemtuzumab ozogamicin (GO) on relapsed/refractory acute promyelocytic leukemia (APL) (2011)
  • Author(s): Takeshita A, et al
  • Organizer: Annual Meeting of American Society of Hematology
  • Place of Presentation: San Diego, USA
• [Presentation] Results of the multi-institutional surveys for the 21st Transfusion Conference of Japanese University Hospitals (2010)
  • Author(s): Fujihara H, Takeshita A, Watanabe H, Suzumura T, Oshida M, Yurugi K, Tomoda Y, Ohto H
  • Organizer: the AABB Annual Meeting and CTTXPO
  • Place of Presentation: Baltimore, Maryland, USA
• [Presentation] Impact of additional chromosomal abnormalities(ACAs) in patients of acute promyelocytic leukemia10years follow up results of the Japan Adult Leukemia Study Group APL97study (2010)
  • Author(s): Ono T, Takeshita A, Kishimoto Y, Kiyoi H, Okada M, Kawai Y, Tsuzuki M, Horikawa K, Matsuda M, Shinagawa K, Monma F, Ohtake S, Nakaseko C, Takahashi M, Yagasaki F, Kimura Y, Iwanaga M, Asou N, Naoe T, Ohno R
  • Organizer: 15th Congress of the European Hematology Association
  • Place of Presentation: Barcelona, Spain
• [Presentation] Clinical Features and Outcomes of Elderly Patients with Acute Promyelocytic Leukemia(APL)-the Japan Adult Leukemia Study Group APL97Study (2010)
  • Author(s): Ono T, Takeshita A, Kishimoto Y, Kiyoi H, Okada M, Kawai Y, Tsuzuki M, Horikawa K, Matsuda M, Shinagawa K, Monma F, Ohtake S, Nakaseko C, Takahashi M, Yagasaki F, Kimura Y, Iwanaga M, Asou N, Naoe T, Ohno R
  • Organizer: 52nd annual meeting of American Society of Hematology
  • Place of Presentation: Orland, FL, USA
• [Presentation] Erythrocyte alloimmunization analyzed by gender and transfusion status (2010)
  • Author(s): Takeshita A, Watanabe H, Fujihara H, Oshida M, Yurugi K, Tomoda Y, Uchikawa M, Kino S, Nollet KE, Ohto H
  • Organizer: 52nd American Society
  • Place of Presentation: Orland, FL, USA
• [Presentation] Erythrocyte allo-immunization analyzed by gender and transfusion status (2010)
  • Author(s): Takeshita A, et al
  • Organizer: 52^<nd> Annual Meeting of American Society of Hematology
  • Place of Presentation: Orange County Convention Center, Orland, Florida
• [Presentation] Clinical features and outcomes in elderly patients with acute promyelocytic leukemia -the Japan Adult Leukemia Study Group APL97 study- (2010)
  • Author(s): Ono T, Takeshita A, et al
  • Organizer: American Society of Hematology (ASH) Annual Meeting and Exposition (52nd)
  • Place of Presentation: Orange County Convention Center, Orlando, Florida
• [Presentation] Collaborative Study Group of Allo-immunity to Antigen Diversity in Asian Populations. Allo-immunity to erythrocytes in relation to blood transfusion and pregnancy ; Cooperative study of allo-immunity to antigen diversity in Asian populations (2009)
  • Author(s): Takeshita A, et al.
  • Organizer: 51th Annual Meeting of the American Society of Hematology
  • Place of Presentation: New Orleans, USA
  • Year and Date: 2009-12-07
• [Presentation] Collaborative study on erythrocyte irregular antibodies in Japan Results from Japanese Study Group of Allo-Immunity to Antigen Diversity in Asian populations (2009)
  • Author(s): Takeshita A, et al.
  • Organizer: 20^<th> Congress of International Society of Blood Transfusion
  • Place of Presentation: Nagoya, Japan
  • Year and Date: 2009-11-15
• [Presentation] Internalization may be important in the efficacy of CMC-544, a new calicheamicin-conjugated anti-CD22 antibody (2009)
  • Author(s): Takeshita A, et al.
  • Organizer: 68^<th> Annual Meeting of the Japanese Cancer Association
  • Place of Presentation: Yokohama, Japan
  • Year and Date: 2009-10-02
• [Presentation] CMC-544, an anti-CD22 immuno-conjugate of calicheamicin, decreases the level of compliment inhibitory factors on malignant B-cell lymphoma cells (2009)
  • Author(s): Takeshita A, et al.
  • Organizer: 14^<th> congress of European Society of Hematology
  • Place of Presentation: Berlin, Germany
  • Year and Date: 2009-06-07
• [Presentation] Internalization speed may be one of the factors in the efficacy of CMC-544(inotuzumab ozogamicin), the newly developed calicheamicin-conjugated antibody (2009)
  • Author(s): Takeshita A, Shinjo K, Watanabe Y, Maekawa M, Ohnishi K, Sugimoto Y, Ohno R
  • Organizer: 68th Annual Meeting of Japanese Cancer Society
  • Place of Presentation: Yokohama
• [Presentation] The role of medical technologists on the education of transfusion medicine : Results of the multi-institutional surveys for the 21th Transfusion Conference of University Hospitals in Japan (2009)
  • Author(s): Watanabe H, Takeshita A, Oshida M, Fujihara H, Tomoda Y, Yurugi K Ohto H
  • Organizer: 20th Congress of International Society of Blood Transfusion
  • Place of Presentation: Nagoya
• [Presentation] Collaborative study on erythrocyte irregular antibodies in Japan Results from Japanese Study Group of Allo-Immunity to Antigen Diversity in Asian populations (2009)
  • Author(s): Takeshita A, Watanabe H, Oshida M, Yurugi K, Tomoda Y, Uchikawa M, Kino S, Ohto H
  • Organizer: 20th Congress of International Society of Blood Transfusion
  • Place of Presentation: Nagoya
• [Presentation] Collaborative Study Group of Allo-immunity to Antigen Diversity in Asian Populations. Differences in allo-immunity to erythrocytes in Asian countries ; Cooperative study of allo-immunity to antigen diversity in Asian populations (2009)
  • Author(s): Watanabe H, Takeshita A, Kim DW, Han KS, Kwon SY, Suh JS, Natalie CPH, Nadarajan VS, Permpikul P, Nuranissa Bte Yacob Marican NBY, Uchikawa M, Kino S, Ohto H
  • Organizer: 51st Annual Meeting of the American Society of Hematology
  • Place of Presentation: New Orleans, LA, USA
• [Presentation] Collaborative Study Group of Allo-immunity to Antigen Diversity in Asian Populations. Allo-immunity to erythrocytes in relation to blood transfusion and pregnancy ; Cooperative study of allo-immunity to antigen diversity in Asian populations (2009)
  • Author(s): Takeshita A, Watanabe H, Kim DW, Han KS, Kwon SY, Suh JS, Natalie CPH, Nadarajan VS, Permpikul P, Nuranissa Bte Yacob Marican NBY, Uchikawa M, Kino S, Ohto H
  • Organizer: 51st Annual Meeting of the American Society of Hematology
  • Place of Presentation: Orleans, LA, USA
• [Presentation] CMC-544, an anti-CD22immunoconjugate of calicheamicin, decreases the level of compliment inhibitory factors on malignant B-cell lymphoma cells (2009)
  • Author(s): Takeshita A, Yamakage N, Shinjo K, Hirano I, Ono T, Fujihara H, Watanabe H, Iino K, Hashimoto D, Maekawa M, Ohnishi K, Ohno R
  • Organizer: 14nd Congress of European Society of Hematology
  • Place of Presentation: Berlin, Germany
• [Presentation] A Phase I/II Trial of Gemtuzumab Ozogamicin in Combination with Cytarabine and Anthracyclines(Daunorubicin or Idarubicin) for CD33-Positive Refractory or Relapsed AML Patients Younger Than65Years. Results of Phase I Parts of the JALSG-AML206Study (2009)
  • Author(s): Usui N, Takeshita A, Nakaseko C, Dobashi N, Fujita H, Kiyoi H, Kobayashi Y, Fujisawa S, Yagasaki F, Nishii K, Kiguchi T, Miyazaki Y, Ohtake S, Miyawaki S, Ohno R, Naoe T, Ohnishi K
  • Organizer: 51st Annual Meeting of the American Society of Hematology
  • Place of Presentation: New Orleans, LA, USA
• [Presentation] Collaborative Study Group of Allo-immunity to Antigen Diversity in Asian Populations : Differences in allo-immunity to erythrocytes in Asian countries ; Cooperative study of allo-immunity to antigen diversity in Asian populations (2009)
  • Author(s): Watanabe H, Takeshita A, Kim DW, Han KS, Kwon SY, Suh JS, Natalie CPH, Nadarajan VS, Permpikul P, Nuranissa Bte Yacob Marican NBY, Uchikawa M, Kino S, Ohto H
  • Organizer: 51st Annual Meeting of the American Society of Hematology
  • Place of Presentation: New Orleans, LA, USA
• [Presentation] The role of medical technologists on the education of transfusion medicine : Results of the multi-institutional surveys for the21th Transfusion Conference of University Hospitals in Japan (2009)
  • Author(s): Watanabe H, Takeshita A, Oshida M, Fujihara H, Tomoda Y, Yurugi K Ohto H
  • Organizer: 20th Congress of International Society of Blood Transfusion
  • Place of Presentation: Nagoya
• [Book] APLにおけるレチノイン酸症候群とケモカイン産生 (2010)
  • Author(s): 竹下明裕
  • Total Pages: 5
  • Publisher: 血液・腫瘍科
• [Book] 血液疾患における分子標的治療 ドラッグラグ解消に向けて 白血病急性前骨髄球性白血病に対するレチノイン酸Am80 最近の進展 (2010)
  • Author(s): 竹下明裕
  • Total Pages: 6
  • Publisher: 血液フロンティア