2003 年度実績報告書

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

研究課題

研究課題/領域番号	14207066
研究機関	慶應義塾大学
研究代表者	吉村泰典慶應義塾大学, 医学部, 教授 (10129736)
研究分担者	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)
キーワード	子宮内膜 / 幹細胞 / 免疫不全マウス / DNAマイクロアレイ
研究概要	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。

研究成果
(37件)

すべてその他

すべて文献書誌 (37件)

[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] Yoshimura Y: "Oocyte maturation in humans : the potential relevance to reproductive medicine."J Mamm Ova Res.. 20. 86-92 (2003)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] Yoshimura Y: "Insulin-like growth factors and their binding proteins : the potential relevance to reproductive physiology."Reprod Med Biol.. 2. 1-24 (2003)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] Matsuzaki Y, Kinjo K, Mulligan RC, Okano H: "Unexpectedly efficient homing capacity of purified murine hematopoietic stem cell."Immunity.. 20. 87-93 (2004)
[文献書誌] Yamashima T, Tonchev BA, Seki T, Sawamoto K, Okano H: "Vascular adventitia generates neuronal progenitors in monkey hippocampus after ischemia."Hippocampus.. (in press). (2004)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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 年度 実績報告書

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

研究代表者

吉村 泰典 慶應義塾大学, 医学部, 教授 (10129736)

研究成果

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

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

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

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

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

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

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

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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 年度実績報告書

吉村泰典慶應義塾大学, 医学部, 教授 (10129736)