膵幹細胞の分化と維持の制御機構

• Project/Area Number: 21390280
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Metabolomics
• Research Institution: Kumamoto University
• Principal Investigator: 粂 昭苑 Kumamoto University, 教授 (70347011)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000); Fiscal Year 2011: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2010: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2009: ¥6,630,000 (Direct Cost: ¥5,100,000、Indirect Cost: ¥1,530,000)
• Keywords: 膵臓 / 肝細胞 / 胚性幹細胞 / 分化誘導 / 増殖

Research Abstract

本研究では、ES/iPS細胞から膵臓系譜への発生途上のモデル細胞、膵臓の体性幹細胞のモデル細胞を基軸とし、正常発生分化・再生過程と常に比較しながら、細胞の内因性プログラムの変危化、分化誘導シグナルの同定、幹細胞の自己複製と分化制御機構(「ニッチ」)解明を目指す。
1.汎腸管内胚葉の細胞表層マーカーとしてのDaf1(CD55)の発見およびその解析
これまでにES細胞由来の内胚葉の解析により、Daf1(CD55)を同定した。CD55+/E-cadherin+細胞はCxcr4+/E-cadherin+に比べて、成熟度の高い細胞集団であることがマイクロアレーおよび細胞培養の結果により強くしさせれた。
2.膵臓の再生・修復における体性幹細胞の同定、遺伝子発現、機能解析
これまでに申請者らが、ES細胞由来のモデル細胞において、Epiplakin1(Eppk1)遺伝子を膵幹細胞マーカー候補遺伝子として同定した(Yoshida T et al.,2008)。Eppk1遺伝子はマウス胎仔Pdxl陽性の膵前駆細胞、Ngn3陽性膵内分泌前駆細胞、Ptfla陽性膵外分泌前駆細胞、成体腺房中心細胞で発現している(Yoshida T et al.,2008)。また、膵障害モデルにおいてEppk1が成体の膵幹細胞の指標となることが強く示唆された。この遺伝子が膵臓のみでなく、肝内胆管で発現しており、肝障害モデルにおいて一過性に現れるOval細胞においても発現することを明らかにした(Matsuo et al.,2011)。Eppk1遺伝子発現を追跡できる, Eppk1-CreERマウスを作成し、レポータマウスとの交配を完了した。今後この遺伝子の発現を追跡することで、肝臓と膵臓の成体前駆細胞を追跡できる。

Research Products

• [Journal Article] Influence of 60 ns Pulsed electric fields on embryonic stem cells. (2011)
  • Author(s): Akiyama, M., Shiraishi E., Sakugawa T, Hosseini, SHR., Akiyama H., Shiraki N., Kume S.
  • Journal Title: IEEE transactions on DEI Volume: (In press)
  • Peer Reviewed
• [Journal Article] Endoderm and mesoderm reciprocal signaling mediated by CXCL12 and CXCR4 regulates the migration of angioblasts and establishes the pancreatic fate. (2011)
  • Author(s): Katsumoto, K., Kume, S.
  • Journal Title: Development Volume: (In press)
  • Peer Reviewed
• [Journal Article] An expression profile analysis of ES cell-derived definitive endodermal cells and Pdx1-expressing cells. (2011)
  • Author(s): Ogaki S, Harada S, Shiraki N, Kume K, Kume S.
  • Journal Title: BMC Developmental Biology Volume: 11 Pages: 13-13
  • Peer Reviewed
• [Journal Article] Epiplakin1 is expressed in the cholangiocyte lineage cells in normal liver and adult progenitor cells in injured liver. (2011)
  • Author(s): Matsuo A., Yoshida T., Yasukawa T., Miki R., Kazuhiko K., Kume S.
  • Journal Title: MOD-Gene Expression Patterns Volume: 11 Pages: 255-262
  • Peer Reviewed
• [Journal Article] In vitro models of pancreatic differentiation using ES or iPS cells. (2011)
  • Author(s): Higuchi Y., Shiraki N., Kume, S
  • Journal Title: Congenital Anomalies Volume: 51 Pages: 21-25
  • Peer Reviewed
• [Journal Article] Xenopus embryos and ES cells as tools for studies of developmental biology. (2011)
  • Author(s): Kume, S.
  • Journal Title: Neurochem Res. Volume: (In press)
  • Peer Reviewed
• [Journal Article] Guiding ES cell differentiation into the definitive endoderm lineages. (2010)
  • Author(s): Shiraki S., Higuchi Y., Kume S
  • Journal Title: Inflamation and Regeneration Volume: 30 Pages: 109-114
  • Peer Reviewed
• [Journal Article] Synthesized basement membranes direct the differentiation of mouse embryonic stem cells into pancreatic lineages. (2010)
  • Author(s): Higuchi Y, Shiraki N, Yamane K, Qin Z, Mochitate K, Hara M, Kume K, Kume S.
  • Journal Title: J.Cell Science Volume: 123 Pages: 2733-2742
  • Peer Reviewed
• [Journal Article] Identification of DAF1/CD55, a novel definitive endoderm marker. (2010)
  • Author(s): Shiraki N, Harada S, Ogaki S, Kume K., Kume S
  • Journal Title: Cell Struct.Funct. Volume: 35 Pages: 73-80
  • Peer Reviewed
• [Journal Article] Embryonic and adult stem cell systems in mammals : Ontology and regulation" in 'Comparative aspects of Stem cells' (2010)
  • Author(s): Katsumoto, K., Shiraki, N., Miki, R., Kume, S.
  • Journal Title: Develop.Growth Diff 52 Pages: 115-129
  • Peer Reviewed
• [Journal Article] Expression of Epiplakin 1 in the developing and adult mouse retina (2010)
  • Author(s): Yoshida, T., Guo, X., Namekata, K., Mitamura, Y., Kume, S., Harada T.
  • Journal Title: Jpn J Ophthalmol 54 Pages: 85-88
  • Peer Reviewed
• [Journal Article] Differentiation and characterization of embryonic stem cells into three germ layers (2009)
  • Author(s): Shiraki, N., Higuchi, Y., Harada, S., Umeda, K., Isagawa, T., Aburatani, H., Kume, K., Kume, S.
  • Journal Title: Biochem.Biophys.Res.Comm. 381 Pages: 694-699
  • Peer Reviewed
• [Journal Article] Analysis of gene expressions of ES-derived Pdx1-expressing cells : Implications of genes involved in pancreas differentiation (2009)
  • Author(s): Yoshida, T., Murata, K., Shiraki, N., Kume, K., Kume, S.
  • Journal Title: Develop.Growth Differ. 51 Pages: 463-472
  • Peer Reviewed
• [Journal Article] Origin of pancreatic precursors and the mechanism of endoderm regionalization in chick embryos (2009)
  • Author(s): Katsumoto, K., Fukuda, K., Kimura, W., Shimamura, K., Yasugi, S., Kume, S.
  • Journal Title: Mech.Dev. 126 Pages: 539-551
  • Peer Reviewed
• [Journal Article] Origin of the ventral pancreatic precursors (2009)
  • Author(s): Matsuura, K., Katsumoto, K., Fukuda, K., Kume, K., Kume, S.
  • Journal Title: Mech.Dev. Pages: 817-827
  • Peer Reviewed
• [Presentation] Epiplakin1 (EPPk1) is expressed in foregut endoderm, also novel marker for adult progenitor cells in injured liver. (2011)
  • Author(s): Akira Matsuo, Tetsu Yoshida, Rika Miki, Kazuhiko Kume, Shoen Kume
  • Organizer: Keystone Symposia on Molecular and cellular biology
  • Place of Presentation: Hilton Santa Fe/Historic Plaza (Santa Fe, New Mexico.)
  • Year and Date: 2011-02-10
• [Presentation] Epiplakin1 (Eppk1) marks the cholangiocytes and transit amplifying cells in injured liver (2010)
  • Author(s): Matsuo A, Yoshida T, Miki R, Fujiwara S, Kume, K., Kume, S.
  • Organizer: 第33回分子生物学会
  • Place of Presentation: 国際会議場〔神戸市〕
  • Year and Date: 2010-12-10
• [Presentation] Analysis of Epiplakin1 expression during pancreatic regeneration (2010)
  • Author(s): Miki R, Yoshida T, Shiraki N, Kume K, Kume S
  • Organizer: 第33回分子生物学会
  • Place of Presentation: 国際会議場〔神戸市〕
  • Year and Date: 2010-12-10
• [Presentation] Regeneration of beta cells in neonatal mouse after streptozotocin treatment (2010)
  • Author(s): Kawamura Y, Miki R, Yasukawa T, Yoshida T, Shiraki N, Kume K, Kume S
  • Organizer: 第33回分子生物学会
  • Place of Presentation: 国際会議場〔神戸市〕
  • Year and Date: 2010-12-10
• [Presentation] The establishment of a screening system for low molecular compounds for β cell inducing activity (2010)
  • Author(s): Sakano, D., Shiraki, N., Kataoka, M., Uesugi, M., Saya, H., Kume, K, Kume, S.
  • Organizer: 第33回分子生物学会
  • Place of Presentation: 国際会議場〔神戸市〕
  • Year and Date: 2010-12-10
• [Presentation] Generation of intestinal epithelial like cell derived from ES cells (2010)
  • Author(s): Ogaki, S., Shiraki, N., Kume, K., Kume, S.
  • Organizer: 第33回分子生物学会
  • Place of Presentation: 国際会議場〔神戸市〕
  • Year and Date: 2010-12-10
• [Presentation] The effects of amino acids on the differentiation of mouse embryonic stem cells into hepatic lineages. (2010)
  • Author(s): Shiraki Y., Shiraki N., Mochida T., Kume S., Endo F
  • Organizer: 第33回分子生物学会
  • Place of Presentation: 国際会議場〔神戸市〕
  • Year and Date: 2010-12-10
• [Presentation] Efficient differentiation of mouse and human ES/iPS cells into hepatic cells using feeder free basement membrane substratum. (2010)
  • Author(s): Shiraki N., Zeng Q., Mochitate K., Higuchi Y., Umeda K., Kume K., Kume S.
  • Organizer: 第33回分子生物学会
  • Place of Presentation: 国際会議場〔神戸市〕
  • Year and Date: 2010-12-10
• [Presentation] A novel culture environment that promotes hepatic differentiation from human iPS and mouse ES cells. (2010)
  • Author(s): Yamazoe, T., Shiraki, N., Umeda, K., Kume, K., Kume, S.
  • Organizer: 第33回分子生物学会
  • Place of Presentation: 国際会議場〔神戸市〕
  • Year and Date: 2010-12-10
• [Presentation] Epiplakin1 (Eppk1) marks the progenitor population in adult liver (2010)
  • Author(s): Matsuo A, Yoshida T, Miki R, Fujiwara S, Kume, K.e, Kume, S.
  • Organizer: The 16th International Conference of the International society of Differentiation
  • Place of Presentation: 市民会館(奈良市)
  • Year and Date: 2010-11-18
• [Presentation] ES/iPS細胞の分化誘導をサポートする細胞外環境の重要性 (2010)
  • Author(s): 粂昭苑
  • Organizer: 専門家と直接意見交換シンポジウム in KRP Part II
  • Place of Presentation: 京都リサーチパーク(京都市)
  • Year and Date: 2010-10-19
• [Presentation] 膵臓・肝臓の再生医療と創薬への応用 (2010)
  • Author(s): 粂昭苑
  • Organizer: Walk Again 2010再生医療シンポジウム
  • Place of Presentation: アクロス福岡・国際会議場(福岡市)
  • Year and Date: 2010-07-25
• [Presentation] Guiding ES/iPS cells to differentiate into pancreatic beta cells (2010)
  • Author(s): 粂昭苑
  • Organizer: 第43回発生生物学会
  • Place of Presentation: 京都国際会議場(京都市)
  • Year and Date: 2010-06-23
• [Presentation] Efficient differentiation of mouse and human ES/iPS cells into hepatic cells using feeder free basement membrane substratum. (2010)
  • Author(s): Shiraki N., Zeng Q., Mochitate K., Higuchi Y., Umeda K., Kume K., Kume S.
  • Organizer: 8^<th> ISSCR Annual Meeting
  • Place of Presentation: MOSCONE CENTER (San Francisco, USA)
  • Year and Date: 2010-06-18
• [Presentation] ES/iPS細胞から膵細胞系譜への分化誘導 (2010)
  • Author(s): 粂昭苑
  • Organizer: 第53回日本糖尿病学会年次学術集会
  • Place of Presentation: 岡山コンベンションセンター(岡山市)
  • Year and Date: 2010-05-27
• [Presentation] 人工基底膜を用いたES細胞から肝細胞への分化誘導 (2010)
  • Author(s): 白木伸明, 樋口裕一郎, 山添太士, 曾勤, 持立克身, 小林直哉, 粂和彦, 粂昭苑
  • Organizer: 第9回日本再生医療学会総会 シンポジウム「肝移植・再生」
  • Place of Presentation: 広島国際会議場(広島市)
  • Year and Date: 2010-03-19
• [Presentation] 幹細胞を用いた消化器官細胞への分化誘導 (2009)
  • Author(s): 粂昭苑
  • Organizer: 第15回大阪大学医療組織工学フォーラム
  • Place of Presentation: 大阪大学(大阪市)
  • Year and Date: 2009-12-01
• [Presentation] 幹細胞を用いた消化器官の発生再生研究 (2009)
  • Author(s): 粂昭苑
  • Organizer: 第33回阿蘇シンポジウム
  • Place of Presentation: 阿蘇リゾートグランヴィリオホテル(阿蘇市)
  • Year and Date: 2009-08-01
• [Presentation] Differentiation of ES cells into pancreatic and hepatic cells (2009)
  • Author(s): 粂昭苑
  • Organizer: ISREC seminar
  • Place of Presentation: Swiss Institute for Experimental Cancer Research(ローザンヌ、スイス連邦)
  • Year and Date: 2009-07-14
• [Presentation] Novel differentiation procedures of the mouse ES or iPS cells into pancreatic cell lineages (2009)
  • Author(s): Yuichiro Higuchi, Keitaro Yamane, Nobuaki Shiraki, Zeng Qin, Katsumi Mochitate, Kazuhiko Kume, Shoen Kume
  • Organizer: 7^<th> ISSCR Annual Meeting
  • Place of Presentation: Centtre Convencion Internacional(バルセロナ、スペイン王国)
  • Year and Date: 2009-07-08
• [Presentation] Efficient differentiation of mouse and human ES cells into hepatic cells using feeder free basement membrane substratum in vitro (2009)
  • Author(s): Nobuaki Shiraki, Zeng Qin, Katsumi Mochitate, Yuichiro Higuchi, Kahoko Umeda, Kazuhiko Kume, Shoen Kume
  • Organizer: 7^<th> ISSCR Annual Meeting
  • Place of Presentation: Centtre Convencion Internacional(バルセロナ、スペイン王国)
  • Year and Date: 2009-07-08
• [Presentation] ES・iPS細胞から消化器官細胞への分化誘導研究 (2009)
  • Author(s): 粂昭苑
  • Organizer: 第49回先天異常学会
  • Place of Presentation: 鹿児島市民文化ホール(鹿児島市)
  • Year and Date: 2009-06-25
• [Presentation] ES・iPS細胞から消化器官細胞への分化誘導研究 (2009)
  • Author(s): 粂昭苑
  • Organizer: 第83回医工学フォーラム
  • Place of Presentation: 京都大学再生医科学研究所(京都市)
  • Year and Date: 2009-05-29
• [Presentation] 幹細胞を用いた消化器官の再生医学研究 (2009)
  • Author(s): 粂昭苑
  • Organizer: 第112回日本小児科学会学術集会
  • Place of Presentation: 奈良県文化会館(奈良市)
  • Year and Date: 2009-04-17
• [Remarks]
  • URL: http://www.imeg.kumamoto-u.ac.jp/divisions/stem_cell_biology/
• [Remarks]
  • URL: http://www.imeg.kumamoto-u.ac.jp/divisions/stem_cell_biology/
• [Patent(Industrial Property Rights)] 多能性幹細胞の分化誘導効率を改善するための方法及び培地 (2010)
  • Inventor(s): 粂昭苑、遠藤文夫、白木伸明、白木恭子、粂和彦
  • Industrial Property Rights Holder: 熊本大学
  • Industrial Property Number: 2010-242774
  • Filing Date: 2010-10-28
• [Patent(Industrial Property Rights)] 腸細胞の製造方法 (2010)
  • Inventor(s): 粂昭苑、白木伸明、粂和彦, 大垣総一郎
  • Industrial Property Rights Holder: 熊本大学
  • Industrial Property Number: 2010-246161
  • Filing Date: 2010-11-02
• [Patent(Industrial Property Rights)] 細胞の分化誘導 (2009)
  • Inventor(s): 粂昭苑, 白木伸明, 樋口裕一郎, 梅田香穂子, 粂和彦, 山根恵太郎, 持立克身
  • Industrial Property Rights Holder: 熊本大学
  • Industrial Property Number: 2009-136520
  • Filing Date: 2009-06-10
• [Patent(Industrial Property Rights)] 内胚葉細胞、腸管細胞膵細胞の検出方法 (2009)
  • Inventor(s): 粂昭苑, 原田聖子, 白木伸明, 粂和彦
  • Industrial Property Rights Holder: 熊本大学
  • Industrial Property Number: 2009-225758
  • Filing Date: 2009-06-30
• [Patent(Industrial Property Rights)] 細胞の分化誘導方法 (2009)
  • Inventor(s): 粂昭苑, 白木伸明, 粂和彦
  • Industrial Property Rights Holder: 熊本大学
  • Industrial Property Number: 2009-254336
  • Filing Date: 2009-11-05