Development of high-speed closed system for analysis and isolation of rare cell populations within the blood

2012 Fiscal Year Annual Research Report

• Project Area: Hyper Bio Assembler for 3D Celluler Innovation
• Project/Area Number: 23106004
• Research Institution: The University of Tokyo
• Principal Investigator: 中内 啓光 東京大学, 医科学研究所, 教授 (40175485)
• Co-Investigator(Kenkyū-buntansha): 金子 新 京都大学, iPS細胞研究所, 准教授 (40361331) ; 江藤 浩之 京都大学, iPS細胞研究所, 教授 (50286986)
• Project Period (FY): 2011-04-01 – 2016-03-31
• Keywords: 微小細胞 / デバイス / がん / 閉鎖系 / 医工学融合

Outline of Annual Research Achievements

本計画研究班では，①現状のフローサイトメトリー機器を更に発展させ“１００万個に一個以下の頻度でしか存在しない細胞（転移がん細胞など）を同定できる”新しい機器の開発を最終目標にしたPOC研究、②輸血システムに必須な各種のデバイス開発、を推進している。①に関連した成果として、平成23年度から継続しているA01班の新井研究室（名古屋大）、金子研究室（阪大）により開発されたマイクロ流路と超高速カメラ撮影システムを用いたマウス骨髄有核細胞、ヒト末梢血有核細胞、ヒト人工多能性幹細胞human induced pluripotent Stem Cell(以下、ヒトiPS細胞)由来赤芽球/赤血球を用いたマイクロ流路通過時間測定を行い、iPS細胞株クローン毎に異なる分化成熟度の差を簡便に高速に測定可能である事を明らかにした。また、②に関しては、A01班　新井研究室（名古屋大）およびA02班　福田研究室（名古屋大）との連携によって、MEMSシステムを応用した血小板産生デバイスのプロトタイプを完成させた。従来のstatic培養条件（培養皿）において、細胞の内的動作（intrinsic 作用）によって巨核球（血小板前駆細胞）から産生された血小板数に比し、本デバイスを用いることで３倍以上の効率で血小板を産生させることに成功した。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

これまで行ってきた２つのテーマ（①末梢血中の超低頻度頻度細胞を同定できる”新しい機器の開発、②輸血システムに必須な各種のデバイス開発）で良好な結果が得られている為。

Strategy for Future Research Activity

多能性幹細胞からin vitroで赤血球への分化を行う際、最後の脱核の過程が非常に難しい。　そこで、今後はこの脱核の過程を効率良く再現できるようなデバイスの開発を試みる。

Research Products

• [Journal Article] Generation of rejuvenated antigen-specific T cells by pluripotency reprogramming and dedifferentiation. (2013)
  • Author(s): Nishimura T, Nakauchi H.(22名、22番目）
  • Journal Title: Cell Stem Cell Volume: 12 Pages: 114-126
  • DOI: doi: 10.1016/j.stem.2012.11.002.
  • Peer Reviewed
• [Journal Article] The early retinal progenitor-expressed gene Sox11 regulates the timing of the differentiation of retinal cells. (2013)
  • Author(s): Usui A, Mochizuki Y, Iida A, Miyauchi E, Satoh S, Sock E, Nakauchi H, Aburatani H, Murakami A, Wegner M, Watanabe S.
  • Journal Title: Development Volume: 140 Pages: 740-50
  • DOI: doi: 10.1002/hep.26293.
  • Peer Reviewed
• [Journal Article] Wnt3a stimulates maturation of impaired neutrophils developed from severe congenital neutropenia patient-derived pluripotent stem cells. (2013)
  • Author(s): Hiramoto T, Ebihara Y, Mizoguchi Y, Nakamura K, Yamaguchi K, Ueno K, Nariai N, Mochizuki S, Yamamoto S, Nagasaki M, Furukawa Y, Tani K, Nakauchi H, Kobayashi M, Tsuji K.
  • Journal Title: Proc Natl Acad Sci U S A Volume: 110 Pages: 3023-8
  • DOI: doi: 10.1073/pnas.1217039110.
  • Peer Reviewed
• [Journal Article] Wnt5a signaling mediates biliary differentiation of fetal hepatic stem/progenitor cells. (2013)
  • Author(s): Kiyohashi K, Nakauchi H, Watanabe M.(19名、18番目）
  • Journal Title: Hepatology Volume: 57 Pages: 印刷中
  • DOI: doi: 10.1002/hep.26293.
  • Peer Reviewed
• [Journal Article] Distinct B-cell lineage commitment distinguishes adult bone marrow hematopoietic stem cells. (2012)
  • Author(s): Ghosn EE, Yamamoto R, Hamanaka S, Yang Y, Herzenberg LA, Nakauchi H, Herzenberg LA.
  • Journal Title: Proc Natl Acad Sci U S A. Volume: 109 Pages: 5394-8
  • DOI: doi: 10.1073/pnas.1121632109.
  • Peer Reviewed
• [Journal Article] Generation of Kidney from Pluripotent Stem Cells via Blastocyst Complementation. (2012)
  • Author(s): Usui J, Kobayashi T, Yamaguchi T, Knisely AS, Nishinakamura R, Nakauchi H.
  • Journal Title: Am J Pathol Volume: 180 Pages: 2417-26
  • DOI: doi: 10.1016/j.ajpath.2012.03.007.
  • Peer Reviewed
• [Journal Article] MT1-MMP plays a critical role in hematopoiesis by regulating HIF-mediated chemokine/cytokine gene transcription within niche cells. (2012)
  • Author(s): Nishida C, Kusubata K, Tashiro Y, Gritli I, Sato A, Ohki-Koizumi M, Morita Y, Nagano M, Sakamoto T, Koshikawa N, Kuchimaru T, Kizaka-Kondoh S, Seiki M, Nakauchi H, Heissig B, Hattori K.
  • Journal Title: Blood Volume: 119 Pages: 5405-16
  • DOI: doi: 10.1182/blood-2011-11-390849.
  • Peer Reviewed
• [Journal Article] Ability of tetraploid rat blastocysts to support fetal development after complementation with embryonic stem cells. (2012)
  • Author(s): Hirabayashi M, Tamura C, Sanbo M, Goto T, Kato-Itoh M, Kobayashi T, Nakauchi H, Hochi S.
  • Journal Title: Mol Reprod Dev Volume: 79 Pages: 402-12
  • DOI: doi: 10.1002/mrd.22043
  • Peer Reviewed
• [Journal Article] Pluripotent stem cells reveal the developmental biology of human megakaryocytes and provide a source of platelets for clinical application. (2012)
  • Author(s): Takayama N, Eto K.
  • Journal Title: Cell Mol Life Sci Volume: 69 Pages: 3419-28
  • DOI: doi: 10.1007/s00018-012-0995-4.
  • Peer Reviewed
• [Journal Article] In vitro generation of megakaryocytes and platelets from human embryonic stem cells and induced pluripotent stem cells. (2012)
  • Author(s): Takayama N, Eto K.
  • Journal Title: Methods Mol Biol Volume: 788 Pages: 205-17
  • DOI: doi: 10.1007/978-1-61779-307-3_15.
  • Peer Reviewed
• [Journal Article] Identification of Hic-5 as a novel regulatory factor for integrin αIIbb3 activation and platelet aggregation in mice. (2012)
  • Author(s): Kim-Kaneyama JR, Miyauchi A, Lei XF, Arita S, Mino T, Takeda N, Kou K, Eto K, Yoshida T, Miyazaki T, Shioda S, Miyazaki A.
  • Journal Title: J Thromb Haemost Volume: 10 Pages: 1867-74
  • DOI: doi: 10.1111/j.1538-7836.2012.04856.x.
  • Peer Reviewed
• [Journal Article] Donor-dependent variations in hepatic differentiation from human-induced pluripotent stem cells. (2012)
  • Author(s): Kajiwara M, Aoi T, Okita K, Takahashi R, Inoue H, Takayama N, Endo H, Eto K, Toguchida J, Uemoto S, Yamanaka S
  • Journal Title: Proc Natl Acad Sci U S A Volume: 109 Pages: 12538-43
  • DOI: doi: 10.1073/pnas.1209979109.
  • Peer Reviewed
• [Journal Article] PKA/CREB Signaling Triggers Initiation of Endothelial and Hematopoietic Cell Differentiation via Etv2 Induction. (2012)
  • Author(s): Yamamizu K, Matsunaga T, Katayama S, Kataoka H, Takayama N, Eto K, Nishikawa S, Yamashita JK.
  • Journal Title: Stem Cells Volume: 30 Pages: 687-96
  • DOI: doi: 10.1002/stem.1041.
  • Peer Reviewed
• [Journal Article] Identification of Rat Rosa26 Locus Enables Generation of Knock-in Rat Lines Ubiquitously Expressing tdTomato. (2012)
  • Author(s): Kobayashi T, Kato-Itoh M, Yamaguchi T, Tamura C, Sanbo M, Hirabayashi M, Nakauchi H.
  • Journal Title: Stem Cells Dev Volume: 21 Pages: 2981-6
  • DOI: doi: 10.1089/scd.2012.0065.
  • Peer Reviewed
• [Journal Article] Highly efficient generation of definitive endoderm lineage from human induced pluripotent stem cells. (2012)
  • Author(s): Sekine K, Takebe T, Suzuki Y, Kamiya A, Nakauchi H, Taniguchi H.
  • Journal Title: Transplant Proc Volume: 44 Pages: 1127-9
  • DOI: doi: 10.1016/j.transproceed.2012.03.001.
  • Peer Reviewed
• [Journal Article] Inhibition of PAI-1 induces neutrophil-driven neoangiogenesis and promotes tissue regeneration via production of angiocrine factors in mice. (2012)
  • Author(s): Tashiro Y, Nishida C, Sato-Kusubata K, Ohki-Koizumi M, Ishihara M, Sato A, Gritli I, Komiyama H, Sato Y, Dan T, Miyata T, Okumura K, Tomiki Y, Sakamoto K, Nakauchi H, Heissig B, Hattori K.
  • Journal Title: Blood Volume: 119 Pages: 6382-93
  • DOI: doi: 10.1182/blood-2011-12-399659.
  • Peer Reviewed
• [Journal Article] Generation of induced pluripotent stem cells from primary chronic myelogenous leukemia patient samples. (2012)
  • Author(s): Kumano K, Arai S, Hosoi M, Taoka K, Takayama N, Otsu M, Nagae G, Ueda K, Nakazaki K, Kamikubo Y, Eto K, Aburatani H, Nakauchi H, Kurokawa M.
  • Journal Title: Blood Volume: 119 Pages: 6234-42
  • DOI: doi: 10.1182/blood-2011-07-367441.
  • Peer Reviewed
• [Journal Article] Bmi1 confers resistance to oxidative stress on hematopoietic stem cells. (2012)
  • Author(s): Nakamura S, Oshima M, Yuan J, Saraya A, Miyagi S, Konuma T, Yamazaki S, Osawa M, Nakauchi H, Koseki H, Iwama A.
  • Journal Title: PLoS One Volume: 7 Pages: e36209
  • DOI: doi: 10.1371/journal.pone.0036209.
  • Peer Reviewed
• [Journal Article] Development of an All-in-One Inducible Lentiviral Vector for Gene Specific Analysis of Reprogramming. (2012)
  • Author(s): Yamaguchi T, Hamanaka S, Kamiya A, Okabe M, Kawarai M, Wakiyama Y, Umino A, Hayama T, Sato H, Lee YS, Kato-Itoh M, Masaki H, Kobayashi T, Yamazaki S, Nakauchi H.
  • Journal Title: PLoS One Volume: 7 Pages: e41007
  • DOI: doi: 10.1371/journal.pone.0041007.
  • Peer Reviewed
• [Journal Article] A retrospective analysis of germline competence in rat embryonic stem cell lines. (2012)
  • Author(s): Hirabayashi M, Tamura C, Sanbo M, Kato-Itoh M, Kobayashi T, Nakauchi H, Hochi S.
  • Journal Title: Transgenic Res Volume: 22 Pages: 411-6
  • DOI: doi: 10.1007/s11248-012-9638-7.
  • Peer Reviewed
• [Journal Article] Enrichment and clonal culture of hepatic stem/progenitor cells during mouse liver development. (2012)
  • Author(s): Kamiya A, Nakauchi H.
  • Journal Title: Methods Mol Biol Volume: 945 Pages: 273-86
  • DOI: doi: 10.1007/978-1-62703-125-7_16.
  • Peer Reviewed
• [Journal Article] Rapid T-cell chimerism switch and memory T-cell expansion are associated with pre-engraftment immune reaction early after cord blood transplantation. (2012)
  • Author(s): Matsuno N, Yamamoto H, Watanabe N, Uchida N, Ota H, Nishida A, Ikebe T, Ishiwata K, Nakano N, Tsuji M, Asano-Mori Y, Izutsu K, Masuoka K, Wake A, Yoneyama A, Nakauchi H, Taniguchi S.
  • Journal Title: Br J Haematol Volume: 160 Pages: 255-8
  • DOI: doi: 10.1111/bjh.12097.
  • Peer Reviewed
• [Journal Article] Stem cell therapy: an exercise in patience and prudence. Stem cell therapy: an exercise in patience and prudence. (2012)
  • Author(s): Lin HT, Otsu M, Nakauchi H.
  • Journal Title: Philos Trans R Soc Lond B Biol Sci Volume: 368 Pages: 20110334
  • DOI: doi: 10.1098/rstb.2011.0334.
  • Peer Reviewed
• [Journal Article] Role of SOX17 in hematopoietic development from human embryonic stem cells. (2012)
  • Author(s): Nakajima-Takagi Y, Osawa M, Oshima M, Takagi H, Miyagi S, Endoh M, Endo TA, Takayama N, Eto K, Toyoda T, Koseki H, Nakauchi H, Iwama A.
  • Journal Title: Blood Volume: 121 Pages: 447-58
  • DOI: doi: 10.1182/blood-2012-05-431403.
  • Peer Reviewed
• [Journal Article] SALL4, a stem cell biomarker in liver cancers. (2012)
  • Author(s): Oikawa T, Kamiya A, Zeniya M, Chikada H, Hyuck AD, Yamazaki Y, Wauthier E, Tajiri H, Miller LD, Wang XW, Reid LM, Nakauchi H.
  • Journal Title: Hepatology Volume: 57 Pages: 1469-83
  • DOI: doi: 10.1002/hep.26159.
  • Peer Reviewed
• [Presentation] In Vivo Clonal Analysis of Hematopoietic Stem Cells Unveils Novel Myeloid Bypass Differentiation Pathways. (2013)
  • Author(s): Hiromitsu Nakauchi
  • Organizer: 2013 USA-Japan Science Conference
  • Place of Presentation: Hawaii USA
  • Year and Date: 2013-03-24 – 2013-03-27
  • Invited
• [Presentation] Non-myelinating Schwann cells in the mouse bone marrow niche maintain hematopoietic stem cell hibernation through TGF-β signaling. (2012)
  • Author(s): Hiromitsu Nakauchi
  • Organizer: Cold Spring Harbor Asia Conferences
  • Place of Presentation: Suzhou, China
  • Year and Date: 2012-12-03 – 2012-12-05
  • Invited
• [Presentation] Glial cell regulation of hematopoietic stem cell hibernation in the bone marrow niche. (2012)
  • Author(s): Hiromitsu Nakauchi
  • Organizer: ESGCT/SFTCG Congress
  • Place of Presentation: Versailles, France
  • Year and Date: 2012-10-27 – 2012-10-31
  • Invited
• [Presentation] Hibernation of Hematopoietic Stem Cells in the Bone Marrow Niche. (2012)
  • Author(s): Hiromitsu Nakauchi
  • Organizer: The University of Edinburgh
  • Place of Presentation: Edinburgh, England
  • Year and Date: 2012-09-05 – 2012-09-05
  • Invited
• [Presentation] Stem Cell Niche and TGF-beta signaling. (2012)
  • Author(s): Hiromitsu Nakauchi
  • Organizer: EMBL Conference, Stem Cells in Cancer and Regenerative Medicine
  • Place of Presentation: Heidelberg, Germany
  • Year and Date: 2012-08-29 – 2012-09-01
  • Invited
• [Presentation] iPS technology and its potential for future medicine. (2012)
  • Author(s): Hiromitsu Nakauchi
  • Organizer: The Babraham Institute
  • Place of Presentation: Cambridge, England
  • Year and Date: 2012-08-20 – 2012-08-20
  • Invited
• [Presentation] Stem cell biology and its potentials for future medicine. (2012)
  • Author(s): Hiromitsu Nakauchi
  • Organizer: University of Zurich
  • Place of Presentation: Zurich, Switzerland
  • Year and Date: 2012-08-16 – 2012-08-16
  • Invited
• [Presentation] iPS technology and its potential for future medicine. (2012)
  • Author(s): Hiromitsu Nakauchi
  • Organizer: University of Geneva Hospital
  • Place of Presentation: Geneva, Switzerland
  • Year and Date: 2012-08-14 – 2012-08-14
  • Invited
• [Presentation] iPS technology and its future therapeutic potentials. (2012)
  • Author(s): Hiromitsu Nakauchi
  • Organizer: The Sanger Institute
  • Place of Presentation: Cambridge, England
  • Year and Date: 2012-07-24 – 2012-07-24
  • Invited
• [Presentation] Generation of functional organs from pluripotent stem cells. (2012)
  • Author(s): Hiromitsu Nakauchi
  • Organizer: Lecture Theatre, Department of Biochemistry, Cambridge Stem Cell Club
  • Place of Presentation: Cambridge, England
  • Year and Date: 2012-07-12 – 2012-07-12
  • Invited
• [Presentation] Generation of Functional organs from pluripotent stem cells: Toward the next generation of regenerative medicine. (2012)
  • Author(s): Hiromitsu Nakauchi
  • Organizer: ISSCR 10th Annual Meeting
  • Place of Presentation: 横浜、日本
  • Year and Date: 2012-06-13 – 2012-06-16
  • Invited
• [Presentation] Isolation, Clonal Characterization and Hibernation of Hematopoietic Stem Cells. (2012)
  • Author(s): Hiromitsu Nakauchi
  • Organizer: Wellcome Trust Centre for Stem Cell Research, University of Cambridge
  • Place of Presentation: Cambridge, England
  • Year and Date: 2012-05-24 – 2012-05-24
  • Invited
• [Presentation] iPS technology and its future therapeutic potentials. (2012)
  • Author(s): Hiromitsu Nakauchi
  • Organizer: The 5th Dutch Society for Stem Cell Research Meeting (DSSCR)
  • Place of Presentation: Amsterdam, Holland
  • Year and Date: 2012-04-11 – 2012-04-14
  • Invited