1999 Fiscal Year Annual Research Report

臍帯血造血幹細胞制御の分子機構の解明とその無血清体外大量培養法への応用

Research Project

Project/Area Number	10470174
Research Institution	The University of Tokyo
Principal Investigator	辻浩一郎東京大学, 医科学研究所, 助教授 (50179991)
Keywords	臍帯血 / 造血幹細胞 / 造血前駆細胞 / NOD / SCIDマウス / 体外増幅
Research Abstract	(目的)臍帯血造血幹細胞の体外大量培養のための条件を検討する。 (方法)臍帯血CD34+細胞を、種々のサイトカインの組合せで培養した後、培養中の造血前駆細胞や造血幹細胞の数を、クローナル培養法やNOD/SCIDマウス移植系を用いて検討した。 (結果)臍帯血CD34+細胞を、stem cell factor(SCF)、Flk2/Flt3 ligand (FL)、thrombopoietin(TPO)、interleukin(IL)-6、IL-6/soluble IL-6 receptor(IL-6/sIL-6R)それぞれ単独で一週間培養した後、NOD/SCIDマウスに移植したところ、全く生着は認められなかった。2種類のサイトカインの組合せでは、SCFとFLの組合せのみが造血幹細胞を維持することができた。そこで、SCF+FL、SCF+FL+TPO、SCF+FL+TPO+IL-6/sIL-6Rの3者の組合せで培養し、造血前駆細胞/幹細胞の増幅を検討した。それぞれの組合せで培養すると、培養細胞数は7.2倍、17.3倍、45.4倍に増加した。クロナール培養法で算定された造血前駆細胞数は、SCF+FLの組合せではほとんど増加しなかったが、SCF+FL+TPOの組合せでは3.1倍、SCF+FL+TPO+IL-6/sIL-6Rでは6.1倍に増加した。さらに、1-2x10^4個の臍帯血CD34+細胞3者の条件下培養後のNOD/SCIDマウスに移植したところ、非培養群では25匹中6匹(24.0%)しか生着が認められなかったが、SCF+FL群では16匹中7匹(43.8%)、SCF+FL+TPO群では14匹中7匹(50.0%)、SCF+FL+TPO+IL-6/sIL-6R群では16匹中13匹(81.2%)で生着が確認された。生着が認められたNOD/SCIDマウスの移植後12週の骨髄細胞におけるヒトCD45+細胞の比率は、非培養群で1.4%、SCF+FL群で2.3%、SCF+FL+TPO群で6.7%、SCF+FL+TPO+IL-6/sIL-6R群で11.5%であったが、それぞれのCD45+細胞の間で各血球系マーカーの発現に大きな差違は認められなかった。 (結論)SCF、FL、TPO、IL-6/sIL-6Rの組み合わせた培養法は、臍帯血造血幹細胞の有効な増幅法と考えられた。

Research Products
(17 results)

All Other

All Publications (17 results)

[Publications] Sui X,Tsuji K,Ebihara Y,Kishimoto T.Nakahara T: "Soluble IL-6 receptor with IL-6 stimulates megakaryopoiesis from human CD34^+ cells through gp130 signaling"Blood. 93. 2525-2532 (1999)
[Publications] Honda H,Tsuji K,Yazaki Y,Hirai H: "Expression of E2A-HLF chimertic protein induced T-cell apoptosis,B-cell maturation arrest,and development of acute lymp"Blood. 93. 2780-2790 (1999)
[Publications] Kinoshita T,Tsuji K,Nakahara T,Miyajima A: "Hepatic differentiation induced by oncostatin M attenuates fetal liver hematopoiesis"Proc Natl Acad Sci USA. 96. 7265-7270 (1999)
[Publications] Yang F,Tsuji K,Watanabe S,: "Differential effects of human granulocyte colony-stimulating factor(hG-CSF) and thrombopoietin on megakaryopoiesis and platelet function in hG-CSF receptor-transgenic mice"Blood. 94. 950-958 (1999)
[Publications] Matsumoto T,Tsuji K,Nakahara T,Yoshimura A: "Suppression of STAT5 functions in liver,mammary glands, and T cells in cytokine-inducible SH2-containing protein 1 transgenic mice"Mol Cell Biol. 19. 6396-6407 (1999)
[Publications] Ishii T,Tsuji K,Nakahata T,Maekawa T: "Expression of stomal cell-derived factor-1/pre-B cell growth -stimulating factor receptor,CXC chemokine receptor 4,on CD34^+ human bone marrow cells is a phenotypic alteration for committed lymphoid progenitors"J Immunol. 163. 3612-3620 (1999)
[Publications] Matsuoka S,Tsuji K,Kikuchi A,Nakahara T: "CD34 expression on long-term repopulating hematopoietic stem cells changes during developmental stages"Blood. (in press).
[Publications] Ebihara Y,Xu M-j Nakahara T,Tsuji K: "Exclusive expression of G-CSF receptor on myeloid progenitor in bone marrow CD34^+cells"Br J Haematol. (in press).
[Publications] Hayashi Y,Honma Y,Tsuji K,Nakahata T: "SN-1,a novel leukemic cell line with t(11;16)(q23;p13): myeloid characteristics and resistance to retinoids and vitamin D3"Cancer Res. (in press).
[Publications] Ueda Y,Tsuji K,Tanaka R,Kobayashi K: "Expansion of human NOD/SCID-repopulating cells by a combination of stem cell factor,Flk2/Flt3 ligand,thrombopoiet receptor"J Clin Invest. (in press).
[Publications] Ueda T,Manabe A,Tsuji K,: "Massive pericardial and pleural effusion with anasarca following allogeneic bone marrow transplantation"Int J Hematol. (in press).
[Publications] Sugiyama H,Tsuji K,Nakahata T: "Peroxisome proliferator-activated receptor are expressed in mouse bone marrow-derived mast cells"FEBS Lett. (in press).
[Publications] Xu M,Matsuoka S,Nakahata T,Tsuji K: "Evidence for presence of primitive megakaryopoiesis in the early yolk sac"Blood. (in press).
[Publications] Nakahata T,Toru H,Tsuji K: "Cytokine regulation of human mast cells"Marone G,Lichtenstein LM,Galli SJ(eds),Academic Press,. (in press).
[Publications] 辻浩一郎: "血液内科学、造血調節機構と造血微小環境"浅野茂隆編、中外医学社(東京). 6 (1999)
[Publications] 辻浩一郎: "内科学教科書、造血幹細胞の性状"黒川清、松澤祐次編、文光堂(東京). 3 (1999)
[Publications] 辻浩一郎: "造血器疾患治療マニュアル、造血器疾患の幹細胞移植"浅野茂隆、池田康夫、内田卓編、メディカルレビュー社(印刷中).

1999 Fiscal Year Annual Research Report

臍帯血造血幹細胞制御の分子機構の解明とその無血清体外大量培養法への応用

Principal Investigator

辻 浩一郎 東京大学, 医科学研究所, 助教授 (50179991)

Research Products

[Publications] Sui X,Tsuji K,Ebihara Y,Kishimoto T.Nakahara T: "Soluble IL-6 receptor with IL-6 stimulates megakaryopoiesis from human CD34^+ cells through gp130 signaling"Blood. 93. 2525-2532 (1999)

[Publications] Honda H,Tsuji K,Yazaki Y,Hirai H: "Expression of E2A-HLF chimertic protein induced T-cell apoptosis,B-cell maturation arrest,and development of acute lymp"Blood. 93. 2780-2790 (1999)

[Publications] Kinoshita T,Tsuji K,Nakahara T,Miyajima A: "Hepatic differentiation induced by oncostatin M attenuates fetal liver hematopoiesis"Proc Natl Acad Sci USA. 96. 7265-7270 (1999)

[Publications] Yang F,Tsuji K,Watanabe S,: "Differential effects of human granulocyte colony-stimulating factor(hG-CSF) and thrombopoietin on megakaryopoiesis and platelet function in hG-CSF receptor-transgenic mice"Blood. 94. 950-958 (1999)

[Publications] Matsumoto T,Tsuji K,Nakahara T,Yoshimura A: "Suppression of STAT5 functions in liver,mammary glands, and T cells in cytokine-inducible SH2-containing protein 1 transgenic mice"Mol Cell Biol. 19. 6396-6407 (1999)

[Publications] Ishii T,Tsuji K,Nakahata T,Maekawa T: "Expression of stomal cell-derived factor-1/pre-B cell growth -stimulating factor receptor,CXC chemokine receptor 4,on CD34^+ human bone marrow cells is a phenotypic alteration for committed lymphoid progenitors"J Immunol. 163. 3612-3620 (1999)

[Publications] Matsuoka S,Tsuji K,Kikuchi A,Nakahara T: "CD34 expression on long-term repopulating hematopoietic stem cells changes during developmental stages"Blood. (in press).

[Publications] Ebihara Y,Xu M-j Nakahara T,Tsuji K: "Exclusive expression of G-CSF receptor on myeloid progenitor in bone marrow CD34^+cells"Br J Haematol. (in press).

[Publications] Hayashi Y,Honma Y,Tsuji K,Nakahata T: "SN-1,a novel leukemic cell line with t(11;16)(q23;p13): myeloid characteristics and resistance to retinoids and vitamin D3"Cancer Res. (in press).

[Publications] Ueda Y,Tsuji K,Tanaka R,Kobayashi K: "Expansion of human NOD/SCID-repopulating cells by a combination of stem cell factor,Flk2/Flt3 ligand,thrombopoiet receptor"J Clin Invest. (in press).

[Publications] Ueda T,Manabe A,Tsuji K,: "Massive pericardial and pleural effusion with anasarca following allogeneic bone marrow transplantation"Int J Hematol. (in press).

[Publications] Sugiyama H,Tsuji K,Nakahata T: "Peroxisome proliferator-activated receptor are expressed in mouse bone marrow-derived mast cells"FEBS Lett. (in press).

[Publications] Xu M,Matsuoka S,Nakahata T,Tsuji K: "Evidence for presence of primitive megakaryopoiesis in the early yolk sac"Blood. (in press).

[Publications] Nakahata T,Toru H,Tsuji K: "Cytokine regulation of human mast cells"Marone G,Lichtenstein LM,Galli SJ(eds),Academic Press,. (in press).

[Publications] 辻浩一郎: "血液内科学、造血調節機構と造血微小環境"浅野茂隆編、中外医学社(東京). 6 (1999)

[Publications] 辻浩一郎: "内科学教科書、造血幹細胞の性状"黒川清、松澤祐次編、文光堂(東京). 3 (1999)

[Publications] 辻浩一郎: "造血器疾患治療マニュアル、造血器疾患の幹細胞移植"浅野茂隆、池田康夫、内田卓編、メディカルレビュー社(印刷中).

辻浩一郎東京大学, 医科学研究所, 助教授 (50179991)