循環極少数細胞を標的とする閉鎖系高速細胞解析分離装置の開発

2013 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)
• Project Period (FY): 2011-07-25 – 2016-03-31
• Keywords: 微小細胞 / デバイス / がん / 閉鎖系 / 医工学融合

Research Abstract

本計画研究班では、現状のフローサイトメトリー機器を更に発展させ生体内において極少数の頻度でしか存在しない細胞（転移がん細胞など）を同定できうる、新しい機器の開発を最終目標にしている。また、これらの技術を応用した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。成果としては、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。さらに、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）との連携研究では、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血小板を産生させることに成功した。今後はより効率かつ迅速な血小板産生が期待される。

Current Status of Research Progress

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

Reason

現在まで行ってきた、成体内に極少数に存在する細胞の同定方法や新しい輸血システムに必須なデバイス開発に興味深い結果を得られている。

Strategy for Future Research Activity

iPS細胞のような細胞から得られた血芽球から脱核を誘導するようなデバイスを開発する。その結果からより効率的なシステムを構築する。

Research Products

• [Journal Article] Histone demethylase Jmjd3 is required for the development of subsets of retinal bipolar cells. (2014)
  • Author(s): Iida A, Iwagawa T, Kuribayashi H, Satoh S, Mochizuki Y, Baba Y, Nakauchi H, Furukawa T, Koseki H, Murakami A, Watanabe S.
  • Journal Title: Proc Natl Acad Sci U S A. Volume: 111 Pages: 3751-6
  • DOI: 10.1073/pnas.1311480111. Epub 2014 Feb 26.
  • Peer Reviewed
• [Journal Article] Nov/CCN3 regulates long-term repopulating activity of murine hematopoietic stem cells via integrin αvβ3. (2014)
  • Author(s): Ishihara J, Umemoto T, Yamato M, Shiratsuchi Y, Takaki S, Petrich BG, Nakauchi H, Eto K, Kitamura T, Okano T.
  • Journal Title: Int J Hematol. Volume: 99 Pages: 393-406
  • DOI: 10.1007/s12185-014-1534-x. Epub 2014 Feb 22.
  • Peer Reviewed
• [Journal Article] Haploinsufficiency of Sf3b1 leads to compromised stem cell function but not to myelodysplasia. (2014)
  • Author(s): Matsunawa M, Yamamoto R, Sanada M, Sato-Otsubo A, Shiozawa Y, Otsu M, Isono K, Koseki H, Nakauchi H, Ogawa S.
  • Journal Title: Leukemia. Volume: 73 Pages: 1-7
  • DOI: 10.1038/leu.2014.73.
  • Peer Reviewed
• [Journal Article] Expandable Megakaryocyte Cell Lines Enable Clinically Applicable Generation of Platelets from Human Induced Pluripotent Stem Cells. (2014)
  • Author(s): Nakamura S, Takayama N, Hirata S, Seo H, Endo H, Ochi K, Fujita KI, Koike T, Harimoto KI, Dohda T, Watanabe A, Okita K, Takahashi N, Sawaguchi A, Yamanaka S, Nakauchi H, Nishimura S, Eto K.
  • Journal Title: Cell Stem Cell. Volume: 14 Pages: 535-48
  • DOI: 10.1016/j.stem.2014.01.011.
  • Peer Reviewed
• [Journal Article] The TIF1β-HP1 System Maintains Transcriptional Integrity of Hematopoietic Stem Cells. (2014)
  • Author(s): Miyagi S, Koide S, Saraya A, Wendt GR, Oshima M, Konuma T, Yamazaki S, Mochizuki-Kashio M, Nakajima-Takagi Y, Wang C, Chiba T, Kitabayashi I, Nakauchi H, Iwama A.
  • Journal Title: Stem Cell Reports. Volume: 2 Pages: 145-52
  • DOI: 10.1016/j.stemcr.2013.12.008. eCollection
  • Peer Reviewed
• [Journal Article] Stage-specific roles for Cxcr4 signaling in murine hematopoietic stem/progenitor cells in the process of bone marrow repopulation. (2014)
  • Author(s): Lai CY, Yamazaki S, Okabe M, Suzuki S, Maeyama Y, Iimura Y, Onodera M, Kakuta S, Iwakura Y, Nojima M, Otsu M, Nakauchi H.
  • Journal Title: Stem Cells. Volume: - Pages: -
  • DOI: 10.1002/stem.1670.
  • Peer Reviewed
• [Journal Article] Homeodomain transcription factor meis1 is a critical regulator of adult bone marrow hematopoiesis. (2014)
  • Author(s): Ariki R, Morikawa S, Mabuchi Y, Suzuki S, Nakatake M, Yoshioka K, Hidano S, Nakauchi H, Matsuzaki Y, Nakamura T, Goitsuka R.
  • Journal Title: PLoS One. Volume: 9 Pages: e87646
  • DOI: 10.1371/journal.pone.0087646. eCollection 2014.
  • Peer Reviewed
• [Journal Article] Use of cell type-specific transcriptome to identify genes specifically involved in Müller glia differentiation during retinal development. (2014)
  • Author(s): Mochizuki Y, Iida A, Lyons E, Kageyama R, Nakauchi H, Murakami A, Watanabe S.
  • Journal Title: Dev Neurobiol. Volume: 74 Pages: 426-37
  • DOI: 10.1002/dneu.22131.
  • Peer Reviewed
• [Journal Article] Enzyme augmentation therapy enhances the therapeutic efficacy of bone marrow transplantation in mucopolysaccharidosis type II mice. (2014)
  • Author(s): Akiyama K, Shimada Y, Higuchi T, Ohtsu M, Nakauchi H, Kobayashi H, Fukuda T, Ida H, Eto Y, Crawford BE, Brown JR, Ohashi T
  • Journal Title: Mol Genet Metab. Volume: 111 Pages: 139-46
  • DOI: 10.1016/j.ymgme.2013.09.013.
  • Peer Reviewed
• [Journal Article] Derivation of Embryonic Stem Cell Lines from Parthenogenetically Developing Rat Blastocysts. (2014)
  • Author(s): Hirabayashi M, Goto T, Tamura C, Sanbo M, Hara H, Kato-Itoh M, Sato H, Kobayashi T, Nakauchi H, Hochi S.
  • Journal Title: Stem Cells Dev. Volume: 23 Pages: 107-14
  • DOI: 10.1089/scd.2013.0200. Epub 2013 Nov 7.
  • Peer Reviewed
• [Journal Article] Immortalization of Erythroblasts by c-MYC and BCL-XL Enables Large-Scale Erythrocyte Production from Human Pluripotent Stem Cells. (2013)
  • Author(s): Hirose S, Takayama N, Nakamura S, Nagasawa K, Ochi K, Hirata S, Yamazaki S, Yamaguchi T, Otsu M, Sano S, Takahashi N, Sawaguchi A, Ito M, Kato T, Nakauchi H, Eto K.
  • Journal Title: Stem Cell Reports. Volume: 1 Pages: 499-508
  • DOI: 10.1016/j.stemcr.2013.10.010.
  • Peer Reviewed
• [Journal Article] Mesenchymal progenitor cells in mouse fetal liver regulate differentiation and proliferation of hepatoblasts. (2013)
  • Author(s): Ito K, Yanagida A, Okada K, Yamazaki Y, Nakauchi H, Kamiya A.
  • Journal Title: Liver Int. Volume: - Pages: -
  • DOI: 10.1111/liv.12387.
  • Peer Reviewed
• [Journal Article] Profiling of MicroRNA in Human and Mouse ES and iPS Cells Reveals Overlapping but Distinct MicroRNA Expression Patterns. (2013)
  • Author(s): Razak SR, Ueno K, Takayama N, Nariai N, Nagasaki M, Saito R, Koso H, Lai CY, Murakami M, Tsuji K, Michiue T, Nakauchi H, Otsu M, Watanabe S.
  • Journal Title: PLoS One. Volume: 8 Pages: e73532
  • DOI: 10.1371/journal.pone.0073532
  • Peer Reviewed
• [Journal Article] Top-down motif engineering of a cytokine receptor for directing ex vivo expansion of hematopoietic stem cells. (2013)
  • Author(s): Saka K, Kawahara M, Teng J, Otsu M, Nakauchi H, Nagamune T.
  • Journal Title: J Biotechnol. Volume: 168 Pages: 659-65
  • DOI: 10.1016/j.jbiotec.2013.09.012.
  • Peer Reviewed
• [Journal Article] Generation of Col2a1-EGFP iPS Cells for Monitoring Chondrogenic Differentiation. (2013)
  • Author(s): Saito T, Yano F, Mori D, Ohba S, Hojo H, Otsu M, Eto K, Nakauchi H, Tanaka S, Chung UI, Kawaguchi H.
  • Journal Title: PLoS One. Volume: 8 Pages: e74137.
  • DOI: 10.1371/journal.pone.0074137.
  • Peer Reviewed
• [Journal Article] Do pluripotent stem cells exist in adult mice as very small embryonic stem cells? (2013)
  • Author(s): Miyanishi M, Mori Y, Seita J, Chen JY, Karten S, Chan CK, Nakauchi H, Weissman IL.
  • Journal Title: Stem Cell Reports. Volume: 1 Pages: 198-208
  • DOI: 10.1016/j.stemcr.2013.07.001.
  • Peer Reviewed
• [Journal Article] Clonal analysis unveils self-renewing lineage-restricted progenitors generated directly from hematopoietic stem cells. (2013)
  • Author(s): Yamamoto R, Morita Y, Ooehara J, Hamanaka S, Onodera M, Rudolph KL, Ema H, Nakauchi H.
  • Journal Title: Cell. Volume: 154 Pages: 1112-26
  • DOI: 10.1016/j.cell.2013.08.007.
  • Peer Reviewed
• [Journal Article] Recurrent mutations in multiple components of the cohesin complex in myeloid neoplasms. (2013)
  • Author(s): Kon A, Shih LY, Minamino M, Sanada M, Shiraishi Y, Nakauchi H, et al.
  • Journal Title: Nature Genet. Volume: 45 Pages: 1232-7
  • DOI: 10.1038/ng.2731. Epub 2013 Aug 18.
  • Peer Reviewed
• [Journal Article] An in vitro expansion system for generation of human iPS cell-derived hepatic progenitor-like cells exhibiting a bipotent differentiation potential. (2013)
  • Author(s): Yanagida A, Ito K, Chikada H, Nakauchi H, Kamiya A.
  • Journal Title: PLoS One. Volume: 8 Pages: e67541.
  • DOI: 10.1371/journal.pone.0067541. Print 2013.
  • Peer Reviewed
• [Journal Article] CAMT-iPS Cells Exhibiting Defective MPL Signaling Dysregulate Megakaryopoiesis and Erythropoiesis. (2013)
  • Author(s): Shinji Hirata, Naoya Takayama,Ryoko Jono-Ohnishi, Hiroshi Endo, Sou Nakamura, Takeaki Dohda, Masanori Nishi, Yuhei Hamazaki, Ei-ichi Ishii, Shin Kaneko, Makoto Otsu, Hiromitsu Nakauchi, Shinji Kunishima, and Koji Eto.
  • Journal Title: J Clin Invest. Volume: 123 Pages: 3802-14
  • DOI: 10.1172/JCI64721.
  • Peer Reviewed
• [Journal Article] Two differential flows in a bioreactor promoted platelet generation from human pluripotent stem cell-derived megakaryocytes. (2013)
  • Author(s): Nakagawa Y, Nakamura S, Nakajima M, Endo H, Dohda T, Takayama N, Nakauchi H, Arai F, Fukuda T, Eto K.
  • Journal Title: Exp Hematology. Volume: 41 Pages: 742-8
  • DOI: 10.1016/j.exphem.2013.04.007. Epub 2013 Apr 22.
  • Peer Reviewed
• [Journal Article] Generation of engraftable hematopoietic stem cells from induced pluripotent stem cells by way of teratoma formation. (2013)
  • Author(s): Suzuki N, Yamazaki S, Yamaguchi T, Okabe M, Masaki H, Takaki S, Otsu M, Nakauchi H.
  • Journal Title: Mol Ther. Volume: 21 Pages: 1424-31
  • DOI: 10.1038/mt.2013.71. Epub 2013 May 14.
  • Peer Reviewed
• [Journal Article] Generation of transgenic mouse line expressing Kusabira Orange throughout body, including erythrocytes, by random segregation of provirus method. (2013)
  • Author(s): Hamanaka S, Ooehara J, Morita Y, Ema H, Takahashi S, Miyawaki A, Otsu M, Yamaguchi T, Onodera M, Nakauchi H.
  • Journal Title: Biochem Biophys Res Commun. Volume: 435 Pages: 586-591
  • DOI: 10.1016/j.bbrc.2013.05.017. Epub 2013 May 17.
  • Peer Reviewed
• [Presentation] "Novel non-stepwise early differentiation pathways of HSCs revealed by 5-lineage in vivo tracing" (2013)
  • Author(s): Hiromitsu Nakauchi
  • Organizer: Gordon Research Conference “Stem Cells & Cancer”
  • Place of Presentation: Les Diablerets, Switzerland
  • Year and Date: 20130421-20130426
  • Invited
• [Presentation] Exploring heterogeneity and hierarchy within the most primitive hematopoietic stem cell compartment
  • Author(s): 中内啓光
  • Organizer: 日本分子生物学会シンポジウム
  • Place of Presentation: 兵庫
  • Invited
• [Book] 幹細胞研究と再生医療 (2013)
  • Author(s): 中内啓光（編集）
  • Total Pages: 238
  • Publisher: 南山堂
• [Book] 直伝！フローサイトメトリー (2013)
  • Author(s): 中内啓光（監修）清田純（編集）
  • Total Pages: 278
  • Publisher: 羊土社