2003 Fiscal Year Annual Research Report

雌性生殖器官における成体幹細胞システムの解明と再生医学への応用

Research Project

Project/Area Number	14207066
Research Institution	Keio University
Principal Investigator	吉村泰典慶應義塾大学, 医学部, 教授 (10129736)
Co-Investigator(Kenkyū-buntansha)	田中守慶應義塾大学, 医学部, 講師 (20207145) 丸山哲夫慶應義塾大学, 医学部, 講師 (10209702) 岡野栄之慶應義塾大学, 医学部, 教授 (60160694) 内田浩慶應義塾大学, 医学部, 助手 (90286534) 松崎有未慶應義塾大学, 医学部, 助手 (50338183)
Keywords	子宮内膜 / 幹細胞 / 免疫不全マウス / DNAマイクロアレイ
Research Abstract	研究年度2年目の平成15年度においては、雌性生殖器官組織のなかで子宮内膜組織に焦点を絞り、その成体幹細胞の同定・分離・培養技術のさらなる確立を初年度に引き続き行った。併せてそれらの細胞生物学的特性の解析に着手した。 1.子宮内膜幹細胞の同定・分離とその内膜組織構築能の検討初年度では、Hoechst33342によるSide Population (SP)分離方法を用いて、子宮内膜の幹細胞様細胞集団を分離し得た。このSP細胞集団はCD31/CD45の発現パターンにより+/+,+/-,-/-の3つの分画に分けられた。免疫不全マウスへのSP細胞移植実験では、構築頻度は低いものの、SP移植部位に腺管構造を有する子宮内膜様組織の再構築が認められた。 2.幹細胞維持および分化のためのin vitro培養系の確立自己複製能と相対的な不死化能の証明のため、各種増殖因子を含んだ培養カクテルやマトリゲルを含めた細胞外基質存在下でSP細胞の長期培養系の確立を目指したが、in vitroの培養は極めて困難であることが判明した。現在、共培養を中心とした新しい培養システムの開発に加えて、さらに様々な培養カクテルを用いてin vitro培養が可能か否かを検証中であり、これは次年度も引き続いて行う。 3.子宮内膜幹細胞特異的発現遺伝子群のプロファイリング内膜SP細胞およびnon-SP細胞よりRNAを抽出し、常法に従い蛍光ラベルした後、GeneChip (Affymetrix社)とハイブリダイズさせ、遺伝子発現プロファイリング解析を行った。現時点の解析結果では、Non-SP細胞に比べてSP細胞に強く発現している遺伝子群にABCG2遺伝子が含まれていた。SP細胞特性を規定するABCG2が強発現していることは、戦略も含めた本実験の妥当性を支持しており、本システムを用いて次年度以降もさらに検討を重ねていく。

Research Products

(37 results)

All Other

All Publications (37 results)

[Publications] Maruvama T, Yamamoto Y, Shimizu A, Masuda H, Sakai N, Sakurai R, Asada H, Yoshimura Y: "Pyrazolo pyrimidine-type inhibitors of src family tyrosine kinases promote ovarian steroid-induced differentiation of human endometrial stromal cells In Vitro."Biol Reprod.. 70(1). 214-221 (2004)
[Publications] Teranishi T, Tanaka M, Kimoto S, Ono Y, Miyakoshi K, Kono T, Yoshimura Y: "Rapid replacement of somatic linker histones with the oocyte-specific linker histone H1 foo in nuclear transfer."Dev Biol.. 266. 76-86 (2004)
[Publications] Matsumoto T, Miyakoshi K, Kasai K, Ishimoto H, Tanaka M, Ikeda K, Yoshimura Y: "Fetal goitrous hypothyroidism followed by neonatal transient hypoerthyroidism."Fetal Diagn Ther.. 18. 459-462 (2003)
[Publications] Yoshimura Y: "Oocyte maturation in humans : the potential relevance to reproductive medicine."J Mamm Ova Res.. 20. 86-92 (2003)
[Publications] Kashiwagi S, Miyakoshi K, Ishimoto H, Tanaka M, Yoshimura Y: "Checkmark fetal heart rate pattern associated with severe fetal hypoxia : A case report."Fetal Diagn.Ther.. 18. 237-239 (2003)
[Publications] Iwahashi K, Kuji N, Fujiwara T, Tanaka H, Takahashi J, Inagaki N, Komatsu S, Yamamoto A, Yoshimura Y, Akagawa K: "Expression of the exocytotic protein syntaxin in mouse oocytes."Reproduction.. 126. 73-81 (2003)
[Publications] Sakai N, Maruyama T, Sakurai R, Masuda H, Yamamoto Y, Shimizu A, Kishi I, Asada H, Yamagoe S, Yoshimura Y: "Involvement of histone acetylation in ovarian steroid-induced decidualization of human endometrial stromal cells."J Biol Chem.. 278(19). 16675-16682 (2003)
[Publications] Tei C, Maruyama T, Kuji N, Miyazaki T, Mikami M, Yoshimura Y: "Reduced expression of αvβ3 integrin in the endometrium of unexplained infertility patients with recurrent IVF-ET failures : Improvement by danazol treatment."J Assist Reprod Genet.. 20(1). 13-20 (2003)
[Publications] Miyakoshi K, Tanaka M, Ueno K, Uehara K, Ishimoto H, Yoshimura Y: "Cutoff value of 1 h, 50 g glucose challenge test for screening of gestational diabetes mellitus in a Japanese population."Diabetes Res Clin Pract.. 60. 63-67 (2003)
[Publications] Tanaka Y, Kato S, Tanaka M, Kuji N, Yoshimura Y: "Structure and expression of the human oocyte-specific histone H1 gene elucidated by direct RT-nested PCR of a single oocyte."Biochem Biophys Res Commun.. 304. 351-357 (2003)
[Publications] Yoshimura Y: "Insulin-like growth factors and their binding proteins : the potential relevance to reproductive physiology."Reprod Med Biol.. 2. 1-24 (2003)
[Publications] Ozawa Y, Nakao K, Takeda J, Akira S, Gruss P, Hirano T, Oguchi Y, Okano H: "Down-regulation of STAT3 activation is required as an intrinsic factor to complete rod photoreceptor differentiation."Mol.Cell.Neurosci.. (in press). (2004)
[Publications] Mikami Y, Okano H, Sakaguchi M, Nakamura M, Shimazaki T, Okano HJ, Kawakami Y, Toyama Y, Toda M: "Implantation of dendritic cells in the injured adult spinal coed results in activation of the endogenous neural/progenitor cells for den novo neurogenesis and axonal regeneration, leading to functional recovery."J.Neurosci Res.. (in press). (2004)
[Publications] Ohba H, Chiyoda T, Endo B, Yano M, Hayakawa Y, Sakaguchi M, Darnell RB, Okano HJ, Okano H: "Sox21 is a repressor of neuronal differentiation and is antagonized by YB-1."Neurosci.Lett.. (in press). (2004)
[Publications] Watanabe K, Nakamura M, Iwanami A, Fujita Y, Kanemura Y, Toyama Y, Okano H: "Comparison between fetal spinal cord-and forebrain-derived neural stem/progenitor cells as a source of transplantation for spinal cord injury."Dev.Neurosci.. (in press). (2004)
[Publications] Ieda M, Fukuda K, Kimura K, Hisaka Y, Kawaguchi H, Shimoda K, Takeshita E, Okano H, Kurihara Y, Kurihara H, Ishida J, Fukamizu A, Salamone L, Howard JF, Ogawa S: "Endothelin-1 regulates cardiac sympathetic nerve innervation in the rodent hearty by controlling nerve growth factor expression."J.Clini.Invest.. (in press). (2004)
[Publications] Murata J, Murayama A, Horii A, Doi K, Harada T, Okano H, Kubo T: "Expression of Musashi1, a neural RNA-binding protein, in the cochler of young adult mice."Neuroscience Letter.. 354. 201-204 (2004)
[Publications] Matsuzaki Y, Kinjo K, Mulligan RC, Okano H: "Unexpectedly efficient homing capacity of purified murine hematopoietic stem cell."Immunity.. 20. 87-93 (2004)
[Publications] Yamashima T, Tonchev BA, Seki T, Sawamoto K, Okano H: "Vascular adventitia generates neuronal progenitors in monkey hippocampus after ischemia."Hippocampus.. (in press). (2004)
[Publications] Okada S, Nakamura M, Mikami Y, Ohsugi Y, Yoshizaki K, Kishimoto T, Toyama Y, Okano H: "Blockade of interleukin-6 receptor ameliorates functional recovery in spinal cord injury."J.Neurosci Res.. (in press). (2004)
[Publications] Hu QD, Ang BT, Karsak M, Hu WP, Cui XY, Duka T, Takeda Y, Chia W, Natesan S, Ng YK, Ling EA, Israel A, Maciag T, Small D, Trifonova R, Kopan R, Okano H, Nakafuku M, Chiba S, Hirai H, Schachner M, Pallen CJ, Watanabe K, Xiao ZC: "F3/Contactin acts as a functional ligand for Notch during oligodendrocyte maturation."Cell.. 115. 163-175 (2003)
[Publications] Yoshida T, Tokunaga A, Nakao, Okano H: "Distinct expression patterns of splicing isoforms of mNumb in the endocrine lineage of developing pancreas."Differentiation.. 70. 486-495 (2003)
[Publications] Miyanomori Y, Kobayashi H, Imai T, Watanabe M, Nagata T, Uesugi S, Okano H, Katahira M: "Origin of figher affinity to RNA of the N-terminal RNA-binding domain than that of the C-terminal one of a mouse neural protein, musashi1, as revealed by comparison of their structures, modes of interaction, surface electrostatic potentials, and backbone dynamics."J.Biol.Chem.. 278. 41309-41315 (2003)
[Publications] Tamaki T, Akatsuka A, Okada, Y, Matsuzaki Y, Okano H, Kimura M: "Growth and differentiation potential of main- and side-population cells derived from murine skeletal muscle."Exp.Cell Res.. 291. 83-90 (2003)
[Publications] Baker H, Kobayashi K, Okano H, Saino-Saito S: "Cortical and striatal expression of tyrosine hydroxylase mRNA in neonatal and adult mice."Cell Mol Neurobiol.. 23. 503-518 (2003)
[Publications] Kanuka H, Kuranaga E, Hiratou T, Igaki T, Nelson B, Okano H, Miura M: "Cytosol-endoplasmic reticulum interplay by Sec61alpha translocon in polyglutamine-mediated neurotoxicity in Drosophila."Proc.Natl.Acad.Sci.USA.. 100. 11723-11728 (2003)
[Publications] Ishizuya-Oka A, Shimizu K, Sakakibara SI, Okano H, Ueda S: "Thyroid hormone-upregulated expression of Musashi-1 is specific for progenitor cells of the adult epithelium during amphibian gastrointestinal remodeling."J Cell Sci.. 116. 3157-3164 (2003)
[Publications] Kokuzawa J, Yoshimura S, Kitajima H, Shinoda J, Kaku Y, Iwama T, Morishita R, Shimazaki T, Okano H, Kunisada T, Sakai N: "Hepatocyte growth factor promotes proliferation and neural differentiation of neural stem cells from mouse embryos."Mol.Cell.Neurosci.. 24. 190-197 (2003)
[Publications] Uchida K, Okano H, Hayashi T, Mine Y, Tanioka Y, Nomura T, Kawase T: "Grafted swine neuroepithelial stem cells can form myelinated axons and both efferent and afferent synapses with xenogeneic rat neurons."J.Neurosci.Res.. 72. 661-669 (2003)
[Publications] Yuasa Y, Okabe M, Yoshikawa S, Tabuchi K, Xiong WC, Hiromi Y, Okano H: "Drosophila homeodomain protein REPO controls glial differentiation by cooperating with ETS and BTB transcription factors."Development.. 130. 2419-2428 (2003)
[Publications] Nakamura Y, Yamamoto M, Oda E, Yamamoto A, Kanemura Y, Hara M, Suzuki A, Yamasaki M, Okano H: "Expression of tubulin beta II in neural stem/progenitor cells and radial fibers during human fetal brain development."Lab Invest.. 83. 479-489 (2003)
[Publications] Sasaki T, Kitagawa K, Sugimura S, Omura-Matsuoka E, Tanaka S, Yagita Y, Okano H, Matsumoto M, Hori M: "Implication of Cyclooxygenase-2 on Enhanced Proliferation of Neural Progenitor Cells in the Adult Mouse Hippocampus After Ischemia."J.Neurosci.Res.. 72. 461-471 (2003)
[Publications] Kayahara T, Sawada M, Takaishi S, Fukui H, Seno H, Fukuzawa H, Suzuki K, Hiai H, Kageyama R, Okano H, Chiba T: "Candidate markers for stem and early progenitor cells, Musashi-1 and Hes1, are expressed in crypt base columnar cells of mouse small intestine."FEBS Letter.. 535. 131-135 (2003)
[Publications] Kuo H-C, Pau FKY, Yeoman RR, Mitalipov SM, Okano H, Wolf DP: "Differentiation of monkey embryonic stem cells into neural lineages."Biology of Reproduction.. 68. 1727-1735 (2003)
[Publications] Tonchev AB, Yamashima T, Zhao L, Okano HJ, Okano H: "Proliferation of neural and neuronal progenitors after global brain ischemia in young adult macaque monkeys."Mol.Cell.Neurosci.. 23. 292-301 (2003)
[Publications] Tonchev AB, Yamashima T, Zhao L, Okano H: "Differential proliferative response in the postischemic hippocampus, temporal cortex and olfactory bulb of adult primates."Glia.. 42. 209-224 (2003)
[Publications] Potten CS, Booth C, Tudor GL, Booth D, Brady G, Hurley P, Ashton G, Clarke G, Sakakibara S, Okano H: "Identification of a putative intestinal stem cell marker and early lineage marker ; Musashi1."Differentiation. 71. 28-41 (2003)

2003 Fiscal Year Annual Research Report

雌性生殖器官における成体幹細胞システムの解明と再生医学への応用

Principal Investigator

吉村 泰典 慶應義塾大学, 医学部, 教授 (10129736)

Research Products

[Publications] Maruvama T, Yamamoto Y, Shimizu A, Masuda H, Sakai N, Sakurai R, Asada H, Yoshimura Y: "Pyrazolo pyrimidine-type inhibitors of src family tyrosine kinases promote ovarian steroid-induced differentiation of human endometrial stromal cells In Vitro."Biol Reprod.. 70(1). 214-221 (2004)

[Publications] Teranishi T, Tanaka M, Kimoto S, Ono Y, Miyakoshi K, Kono T, Yoshimura Y: "Rapid replacement of somatic linker histones with the oocyte-specific linker histone H1 foo in nuclear transfer."Dev Biol.. 266. 76-86 (2004)

[Publications] Matsumoto T, Miyakoshi K, Kasai K, Ishimoto H, Tanaka M, Ikeda K, Yoshimura Y: "Fetal goitrous hypothyroidism followed by neonatal transient hypoerthyroidism."Fetal Diagn Ther.. 18. 459-462 (2003)

[Publications] Yoshimura Y: "Oocyte maturation in humans : the potential relevance to reproductive medicine."J Mamm Ova Res.. 20. 86-92 (2003)

[Publications] Kashiwagi S, Miyakoshi K, Ishimoto H, Tanaka M, Yoshimura Y: "Checkmark fetal heart rate pattern associated with severe fetal hypoxia : A case report."Fetal Diagn.Ther.. 18. 237-239 (2003)

[Publications] Iwahashi K, Kuji N, Fujiwara T, Tanaka H, Takahashi J, Inagaki N, Komatsu S, Yamamoto A, Yoshimura Y, Akagawa K: "Expression of the exocytotic protein syntaxin in mouse oocytes."Reproduction.. 126. 73-81 (2003)

[Publications] Sakai N, Maruyama T, Sakurai R, Masuda H, Yamamoto Y, Shimizu A, Kishi I, Asada H, Yamagoe S, Yoshimura Y: "Involvement of histone acetylation in ovarian steroid-induced decidualization of human endometrial stromal cells."J Biol Chem.. 278(19). 16675-16682 (2003)

[Publications] Tei C, Maruyama T, Kuji N, Miyazaki T, Mikami M, Yoshimura Y: "Reduced expression of αvβ3 integrin in the endometrium of unexplained infertility patients with recurrent IVF-ET failures : Improvement by danazol treatment."J Assist Reprod Genet.. 20(1). 13-20 (2003)

[Publications] Miyakoshi K, Tanaka M, Ueno K, Uehara K, Ishimoto H, Yoshimura Y: "Cutoff value of 1 h, 50 g glucose challenge test for screening of gestational diabetes mellitus in a Japanese population."Diabetes Res Clin Pract.. 60. 63-67 (2003)

[Publications] Tanaka Y, Kato S, Tanaka M, Kuji N, Yoshimura Y: "Structure and expression of the human oocyte-specific histone H1 gene elucidated by direct RT-nested PCR of a single oocyte."Biochem Biophys Res Commun.. 304. 351-357 (2003)

[Publications] Yoshimura Y: "Insulin-like growth factors and their binding proteins : the potential relevance to reproductive physiology."Reprod Med Biol.. 2. 1-24 (2003)

[Publications] Ozawa Y, Nakao K, Takeda J, Akira S, Gruss P, Hirano T, Oguchi Y, Okano H: "Down-regulation of STAT3 activation is required as an intrinsic factor to complete rod photoreceptor differentiation."Mol.Cell.Neurosci.. (in press). (2004)

[Publications] Ohba H, Chiyoda T, Endo B, Yano M, Hayakawa Y, Sakaguchi M, Darnell RB, Okano HJ, Okano H: "Sox21 is a repressor of neuronal differentiation and is antagonized by YB-1."Neurosci.Lett.. (in press). (2004)

[Publications] Watanabe K, Nakamura M, Iwanami A, Fujita Y, Kanemura Y, Toyama Y, Okano H: "Comparison between fetal spinal cord-and forebrain-derived neural stem/progenitor cells as a source of transplantation for spinal cord injury."Dev.Neurosci.. (in press). (2004)

[Publications] Murata J, Murayama A, Horii A, Doi K, Harada T, Okano H, Kubo T: "Expression of Musashi1, a neural RNA-binding protein, in the cochler of young adult mice."Neuroscience Letter.. 354. 201-204 (2004)

[Publications] Matsuzaki Y, Kinjo K, Mulligan RC, Okano H: "Unexpectedly efficient homing capacity of purified murine hematopoietic stem cell."Immunity.. 20. 87-93 (2004)

[Publications] Yamashima T, Tonchev BA, Seki T, Sawamoto K, Okano H: "Vascular adventitia generates neuronal progenitors in monkey hippocampus after ischemia."Hippocampus.. (in press). (2004)

[Publications] Okada S, Nakamura M, Mikami Y, Ohsugi Y, Yoshizaki K, Kishimoto T, Toyama Y, Okano H: "Blockade of interleukin-6 receptor ameliorates functional recovery in spinal cord injury."J.Neurosci Res.. (in press). (2004)

[Publications] Yoshida T, Tokunaga A, Nakao, Okano H: "Distinct expression patterns of splicing isoforms of mNumb in the endocrine lineage of developing pancreas."Differentiation.. 70. 486-495 (2003)

[Publications] Tamaki T, Akatsuka A, Okada, Y, Matsuzaki Y, Okano H, Kimura M: "Growth and differentiation potential of main- and side-population cells derived from murine skeletal muscle."Exp.Cell Res.. 291. 83-90 (2003)

[Publications] Baker H, Kobayashi K, Okano H, Saino-Saito S: "Cortical and striatal expression of tyrosine hydroxylase mRNA in neonatal and adult mice."Cell Mol Neurobiol.. 23. 503-518 (2003)

[Publications] Kanuka H, Kuranaga E, Hiratou T, Igaki T, Nelson B, Okano H, Miura M: "Cytosol-endoplasmic reticulum interplay by Sec61alpha translocon in polyglutamine-mediated neurotoxicity in Drosophila."Proc.Natl.Acad.Sci.USA.. 100. 11723-11728 (2003)

[Publications] Ishizuya-Oka A, Shimizu K, Sakakibara SI, Okano H, Ueda S: "Thyroid hormone-upregulated expression of Musashi-1 is specific for progenitor cells of the adult epithelium during amphibian gastrointestinal remodeling."J Cell Sci.. 116. 3157-3164 (2003)

[Publications] Kokuzawa J, Yoshimura S, Kitajima H, Shinoda J, Kaku Y, Iwama T, Morishita R, Shimazaki T, Okano H, Kunisada T, Sakai N: "Hepatocyte growth factor promotes proliferation and neural differentiation of neural stem cells from mouse embryos."Mol.Cell.Neurosci.. 24. 190-197 (2003)

[Publications] Uchida K, Okano H, Hayashi T, Mine Y, Tanioka Y, Nomura T, Kawase T: "Grafted swine neuroepithelial stem cells can form myelinated axons and both efferent and afferent synapses with xenogeneic rat neurons."J.Neurosci.Res.. 72. 661-669 (2003)

[Publications] Yuasa Y, Okabe M, Yoshikawa S, Tabuchi K, Xiong WC, Hiromi Y, Okano H: "Drosophila homeodomain protein REPO controls glial differentiation by cooperating with ETS and BTB transcription factors."Development.. 130. 2419-2428 (2003)

[Publications] Nakamura Y, Yamamoto M, Oda E, Yamamoto A, Kanemura Y, Hara M, Suzuki A, Yamasaki M, Okano H: "Expression of tubulin beta II in neural stem/progenitor cells and radial fibers during human fetal brain development."Lab Invest.. 83. 479-489 (2003)

[Publications] Sasaki T, Kitagawa K, Sugimura S, Omura-Matsuoka E, Tanaka S, Yagita Y, Okano H, Matsumoto M, Hori M: "Implication of Cyclooxygenase-2 on Enhanced Proliferation of Neural Progenitor Cells in the Adult Mouse Hippocampus After Ischemia."J.Neurosci.Res.. 72. 461-471 (2003)

[Publications] Kuo H-C, Pau FKY, Yeoman RR, Mitalipov SM, Okano H, Wolf DP: "Differentiation of monkey embryonic stem cells into neural lineages."Biology of Reproduction.. 68. 1727-1735 (2003)

[Publications] Tonchev AB, Yamashima T, Zhao L, Okano HJ, Okano H: "Proliferation of neural and neuronal progenitors after global brain ischemia in young adult macaque monkeys."Mol.Cell.Neurosci.. 23. 292-301 (2003)

[Publications] Tonchev AB, Yamashima T, Zhao L, Okano H: "Differential proliferative response in the postischemic hippocampus, temporal cortex and olfactory bulb of adult primates."Glia.. 42. 209-224 (2003)

[Publications] Potten CS, Booth C, Tudor GL, Booth D, Brady G, Hurley P, Ashton G, Clarke G, Sakakibara S, Okano H: "Identification of a putative intestinal stem cell marker and early lineage marker ; Musashi1."Differentiation. 71. 28-41 (2003)

吉村泰典慶應義塾大学, 医学部, 教授 (10129736)