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全胚胎仔培養法を用いたマウス二次造血の発生部位の同定

Research Project

Project/Area Number 12036205
Research Category

Grant-in-Aid for Scientific Research on Priority Areas (A)

Allocation TypeSingle-year Grants
Review Section Biological Sciences
Research InstitutionThe University of Tokyo

Principal Investigator

辻 浩一郎  東京大学, 医科学研究所, 助教授 (50179991)

Project Period (FY) 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2000: ¥2,900,000 (Direct Cost: ¥2,900,000)
Keywords全胚胎仔培養法 / 二次造血 / 卵黄嚢 / B細胞
Research Abstract

(目的)胎仔キメラを作製して、成体まで発育させることが比較的容易な鳥類・両生類において、造血発生研究は大きな進歩を遂げた。しかしほ乳類では、着床後の胎仔を取り出した後成体まで発育させることが困難なため、造血発生研究、特に卵黄嚢造血の解析が遅れている。そこで我々は、卵黄嚢(YS)における造血発生を明らかにする目的で、血流開始前のマウス胎仔でgraft YSとhost YSとのキメラを作製後、全胚胎仔培養法にて培養し、graft YS由来細胞からのB細胞の発生について検討した。
(方法)血流開始前のE8.25で、C57BL/6マウスの胎仔を母胎より分離した。graft YSには、Ly5.1マウス、、グリーンマウスを使用し、血島をすべて含むようにgraft YSを分離した。graft YSを顕微鏡下でhost YSへ植え込み、E11.0まで全胚胎仔培養装置で培養した。graft YSが生着したか否かは血流の有無で判定した。培養後40体節以上のものを選択し、graft YS由来の細胞が、血流中にどの程度存在しているかをフローサイトメトリーにより解析した。また、キメラ胎仔のAGM領域を分離し、OP9細胞との共培養系を用いて、graft YS由来のB細胞の出現を検討した。
(結果と考察)作製したキメラのうち、約半数に血流が認められ、それらの血流中にはgraft YS由来の細胞が12.5%存在した。キメラ胎仔のAGM領域より分離された細胞をOP9細胞との共培養すると、B220+/CD19+B細胞が出現し、Ly5.1マウスキメラ、グリーンマウスキメラ、いずれにおいても、半数でgraft YS由来のB細胞が検出された。以上の結果より、血流開始前のYSにも、B細胞に分化する能力をもった細胞が存在し、これらの細胞は確立された血流を介して胎仔内に移動することが示された。本システムによりマウス卵黄嚢造血を経時的に観察することが可能となった。本システムを用いることにより、マウスにおいても造血幹細胞を含めた造血発生のより詳細な解析が期待される。

Report

(1 results)
  • 2000 Annual Research Report
  • Research Products

    (37 results)

All Other

All Publications (37 results)

  • [Publications] Ueda T.: "Expansion of human NOD/SCID-repopulating cells by a combination of stem cell factor, Flk2/Flt3 ligand, thrombopoietin and a complex of interleukin-6 and soluble interleukin-6 receptor."J.Clin.Invest.. 105. 1013-1021 (2000)

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  • [Publications] Hayashi Y.: "SN-1, a novel leukemic cell line with t (11;16)(q23;p13): myeloid characteristics and resistance to retinoids and vitamin D3."Cancer Res.. 60. 1139-1145 (2000)

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  • [Publications] Ebihara Y: "Exclusive expression of G-CSF receptor on myeloid progenitors in bone marrow CD34^+ cells."Br.J.Haematol.. 109. 153-161 (2000)

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  • [Publications] Ueda T.: "Hematopoietic repopulating ability of cord blood CD34^+ cells in NOD/Shi-scid mice."Stem Cells. 18. 204-213 (2000)

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  • [Publications] Ueda T.: "Massive pericardial and pleural effusion with anasarca following allogeneic bone marrow transplantation."Int.J.Hematol.. 71. 394-397 (2000)

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  • [Publications] Sato T.: "Erythroid progenitors differentiate and mature in response to endogenous erythropoietin."J.Clin.Invest.. 106. 263-270 (2000)

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  • [Publications] Ishii t.: "Improvement in bronchiolitis obliterans organizing pneumonia by combination of oral predonisolone and low dose erythromycin in a child after allogegeneic bone marrow transplantation."Bone Marrow Transplant. 26. 907-910 (2000)

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  • [Publications] Sugiyama H.: "Peroxisome proliferator-activated receptors are expressed in mouse bone marrow-derived mast cells."FEBS Lett.. 467. 259-262 (2000)

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  • [Publications] Yoshino H.: "Natural killer cell depletion by anti-asialo GM1 antiserum treatment enhances human hematopoietic stem cell engraftment in NOD/Shi-scid mice."Bone Marrow Transplant.. 26. 1211-1216 (2000)

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  • [Publications] Takaki S.: "Control of B cell production by the adaptor protein Lnk, a negative regulator of tyrosine kinase signaling."Immunity. 13. 599-609 (2000)

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  • [Publications] Matsui A.: "gp130 and c-kit signals synergistically induce thrombopoietin production by hematopoietic cells."Int.J.Hematol.. 72. 455-462 (2001)

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  • [Publications] Tsuji K.: "Expression of G-CSF receptor on myeloid progemtors."Leukemia and Lymphoma. (in press).

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  • [Publications] Matsuoka S.: "CD34 expression on long-term repopulating hematopoietic stem cells changes during developmental stages."Blood. 97. 419-425 (2001)

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  • [Publications] Ma F: "Cytokine requirement for the development of T-lineage potential from lymphohematopoietic progenitor cells."Br.J.Haematol.. (in press).

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  • [Publications] Sugiyama H.: "Peroxisome proliferator-activated receptors are expressed in human cultured mast cells : a possible role of these receptors in negative regulation of mast cell activation."Eur.J.Immunol.. (in press).

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  • [Publications] Xu M.: "Evidence for the presence of primitive megakaryopoiesis in the early yolk sac."Blood. (in press).

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  • [Publications] Ueda T.: "Hematopoietic capability of cord blood CD34^+ cells : a comparison with that of adult bone marrow CD34^+ cells."Int.J.Haematol.. (in press).

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  • [Publications] Yoshimasu T.: "Prompt and durable hematopoietic reconstitution by an unrelated cord blood transplantation in a child with Fanconi anemia."Bone Marrow Transplant.. (in press).

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  • [Publications] Miyamoto K.: "Inhibitory effect of interleukin-3 on the early development of human B-lymphopoiesis."Br.J.Haematol.. (in press).

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  • [Publications] Matsuoka S.: "Generation of definitive hematopoietic stem cells from murine early yolk sac and paraaortic splanchnopleura by AGM region-derived stromal cells."Blood. (in press).

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  • [Publications] Hisakawa H.: "Human granulocyte-macrophage colony-stimulating factor (hGM-CSF)stimulates primitive and definitive erythropoiesis in mouse embryo expressing hGM-CSF receptors but not erythropoietin receptors."Blood. (in press).

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  • [Publications] Ma F.: "Development of human lymphohematopoietic stem/progenitor cells defined by CD34 and CD81 expressions."Blood. (in press).

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  • [Publications] 石井武文: "急性リンパ性白血病治療後期に発症したL-アスパラギナーゼによる壊死性膵炎"日小血会誌. 14. 328-332 (2000)

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  • [Publications] 岩本彰太郎: "大量rhG-CSF療法中にmonosomy7を伴うMDSに移行したKostmann症候群"日小血会誌. 14. 338-345 (2000)

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  • [Publications] 三井哲夫: "造血幹細胞移植における支持療法と今後の課題"Biotherapy. 14. 292-300 (2000)

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  • [Publications] 辻浩一郎: "増幅造血幹細胞移植ガイドライン"分子細胞療法. 1. 197-201 (2000)

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  • [Publications] 海老原康博: "造血幹細胞の同定検査 2)コロニー法"検査と技術. 28. 971-972 (2000)

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  • [Publications] 辻浩一郎: "造血幹細胞の起源と性状"医学のあゆみ. 194. 1021-1025 (2000)

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  • [Publications] 杉山大介: "AGM領域"分子細胞療法. 1. 92-93 (2000)

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  • [Publications] 辻浩一郎: "造血幹細胞の増幅"血液・免疫・腫瘍. (印刷中).

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  • [Publications] 辻浩一郎: "試験管内造血幹細胞増幅"現代医療. (印刷中).

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  • [Publications] 辻浩一郎: "造血幹細胞コロニー刺激因子と臍帯血造血幹細胞の体外増幅"日本産婦人科・新生児血液学会. (印刷中).

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  • [Publications] Nakahata T.: "Cell Therapy"Springer-Verlag, Tokyo. 2 (2000)

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  • [Publications] Nakahata T.: "Mast Cell Biology"Academic Press (in press).

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  • [Publications] Tsuji K.: "Methods in Molecular Medicine-Cytokines and Colony Stimulating Factors"The Human Press.(in press).

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  • [Publications] 田中竜平: "造血器疾患治療マニュアル"メディカルレビュー社(大阪)(印刷中).

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  • [Publications] 辻浩一郎: "小児の症候群"診断と治療社(東京)(印刷中).

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Published: 2000-04-01   Modified: 2018-03-28  

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