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Article ; Online: Induction of memory-like CD8+ T cells and CD4+ T cells from human naive T cells in culture.

Tokumoto, Yasuhito / Araki, Yasuto / Narizuka, Yusuke / Mizuno, Yosuke / Ohshima, Susumu / Mimura, Toshihide

2022 Volume 207, Issue 1, Page(s) 95–103

Abstract: Memory T cells are crucial players in vertebrate adaptive immunity but their development is incompletely understood. Here, we describe a method to produce human memory-like T cells from naive human T cells in culture. Using commercially available human T- ...

Abstract	Memory T cells are crucial players in vertebrate adaptive immunity but their development is incompletely understood. Here, we describe a method to produce human memory-like T cells from naive human T cells in culture. Using commercially available human T-cell differentiation kits, both purified naive CD8+ T cells and purified naive CD4+ T cells were activated via T-cell receptor signaling and appropriate cytokines for several days in culture. All the T-cell activators were then removed from the medium and the cultures were continued in hypoxic condition (1% O2 atmosphere) for several more days; during this period, most of the cells died, but some survived in a quiescent state for a month. The survivors had small round cell bodies, expressed differentiation markers characteristic of memory T cells and restarted proliferation when the T-cell activators were added back. We could also induce memory-like T cells from naive human T cells without hypoxia, if we froze the activated T cells or prepared the naive T cells from chilled filter buffy coats.
MeSH term(s)	CD4-Positive T-Lymphocytes ; CD8-Positive T-Lymphocytes ; Cell Differentiation ; Humans ; Immunologic Memory ; Lymphocyte Activation
Language	English
Publishing date	2022-01-12
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	218531-3
ISSN	1365-2249 ; 0009-9104 ; 0964-2536
ISSN (online)	1365-2249
ISSN	0009-9104 ; 0964-2536
DOI	10.1093/cei/uxab012
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Article ; Online: Overexpression of cyclin dependent kinase inhibitor P27/Kip1 increases oligodendrocyte differentiation from induced pluripotent stem cells.

Tamaki, Shinpei / Tokumoto, Yasuhito

In vitro cellular & developmental biology. Animal

2014 Volume 50, Issue 8, Page(s) 778–785

Abstract: Cell transplantation therapy with oligodendrocyte precursor cells (OPCs) is a promising and effective treatment for diseases involving demyelination in the central nervous system (CNS). In previous studies, we succeeded in producing O4(+) ... ...

Abstract	Cell transplantation therapy with oligodendrocyte precursor cells (OPCs) is a promising and effective treatment for diseases involving demyelination in the central nervous system (CNS). In previous studies, we succeeded in producing O4(+) oligodendrocytes (OLs) from mouse- and human-induced pluripotent stem cells (iPSCs) in vitro; however, the efficiency of differentiation into OLs was lower for iPSCs than that for embryonic stem cells (ESCs). To clarify the cause of this difference, we compared the expression of proteins that contribute to OL differentiation in mouse iPSC-derived cells and in mouse ESC-derived cells. The results showed that the expression levels of cyclin dependent kinase inhibitor P27/Kip1, mitogen-activated protein kinase (MAPK) JNK3, and transcription factor Mash1 were lower in iPSC-derived cells. In contrast, the expression levels of MAPK P38α, P38γ, and thyroid hormone receptor β1 were higher in iPSC-derived cells. We attempted to compensate for the expression changes in P27/Kip1 protein and Mash1 protein in iPSC-derived cells through retrovirus vector-mediated gene expression. Although the overexpression of Mash1 had no effect, the overexpression of P27/Kip1 increased the differentiation efficiency of iPSC-derived cells into O4(+) OLs.
MeSH term(s)	Animals ; Blotting, Western ; Cell Differentiation/physiology ; Cyclin-Dependent Kinase Inhibitor p27/biosynthesis ; Cyclin-Dependent Kinase Inhibitor p27/metabolism ; Cyclin-Dependent Kinase Inhibitor p27/physiology ; Gene Expression/physiology ; Induced Pluripotent Stem Cells/metabolism ; Induced Pluripotent Stem Cells/physiology ; Mice ; Oligodendroglia/metabolism ; Oligodendroglia/physiology
Chemical Substances	Cdkn1b protein, mouse ; Cyclin-Dependent Kinase Inhibitor p27 (147604-94-2)
Language	English
Publishing date	2014-04-25
Publishing country	Germany
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1077810-x
ISSN	1543-706X ; 0883-8364 ; 1071-2690
ISSN (online)	1543-706X
ISSN	0883-8364 ; 1071-2690
DOI	10.1007/s11626-014-9753-2
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Article: Overexpression of cyclin dependent kinase inhibitor P27/Kip1 increases oligodendrocyte differentiation from induced pluripotent stem cells

Tamaki, Shinpei / Tokumoto, Yasuhito

In vitro cellular & developmental biology. 2014 Sept., v. 50, no. 8

2014

Abstract	Cell transplantation therapy with oligodendrocyte precursor cells (OPCs) is a promising and effective treatment for diseases involving demyelination in the central nervous system (CNS). In previous studies, we succeeded in producing O4⁺ oligodendrocytes (OLs) from mouse- and human-induced pluripotent stem cells (iPSCs) in vitro; however, the efficiency of differentiation into OLs was lower for iPSCs than that for embryonic stem cells (ESCs). To clarify the cause of this difference, we compared the expression of proteins that contribute to OL differentiation in mouse iPSC-derived cells and in mouse ESC-derived cells. The results showed that the expression levels of cyclin dependent kinase inhibitor P27/Kip1, mitogen-activated protein kinase (MAPK) JNK3, and transcription factor Mash1 were lower in iPSC-derived cells. In contrast, the expression levels of MAPK P38α, P38γ, and thyroid hormone receptor β1 were higher in iPSC-derived cells. We attempted to compensate for the expression changes in P27/Kip1 protein and Mash1 protein in iPSC-derived cells through retrovirus vector-mediated gene expression. Although the overexpression of Mash1 had no effect, the overexpression of P27/Kip1 increased the differentiation efficiency of iPSC-derived cells into O4⁺ OLs.
Keywords	cell transplantation ; central nervous system ; embryonic stem cells ; gene expression ; induced pluripotent stem cells ; mice ; mitogen-activated protein kinase ; thyroid hormone receptors ; transcription factors
Language	English
Dates of publication	2014-09
Size	p. 778-785.
Publishing place	Springer-Verlag
Document type	Article
ISSN	1071-2690
DOI	10.1007/s11626-014-9753-2
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Quiescence of adult oligodendrocyte precursor cells requires thyroid hormone and hypoxia to activate Runx1.

Tokumoto, Yasuhito / Tamaki, Shinpei / Kabe, Yasuaki / Takubo, Keiyo / Suematsu, Makoto

Scientific reports

2017 Volume 7, Issue 1, Page(s) 1019

Abstract: The adult mammalian central nervous system (CNS) contains a population of slowly dividing oligodendrocyte precursor cells (OPCs), i.e., adult OPCs, which supply new oligodendrocytes throughout the life of animal. While adult OPCs develop from rapidly ... ...

Abstract	The adult mammalian central nervous system (CNS) contains a population of slowly dividing oligodendrocyte precursor cells (OPCs), i.e., adult OPCs, which supply new oligodendrocytes throughout the life of animal. While adult OPCs develop from rapidly dividing perinatal OPCs, the mechanisms underlying their quiescence remain unknown. Here, we show that perinatal rodent OPCs cultured with thyroid hormone (TH) under hypoxia become quiescent and acquire adult OPCs-like characteristics. The cyclin-dependent kinase inhibitor p15/INK4b plays crucial roles in the TH-dependent cell cycle deceleration in OPCs under hypoxia. Klf9 is a direct target of TH-dependent signaling. Under hypoxic conditions, hypoxia-inducible factors mediates runt-related transcription factor 1 activity to induce G1 arrest in OPCs through enhancing TH-dependent p15/INK4b expression. As adult OPCs display phenotypes of adult somatic stem cells in the CNS, the current results shed light on environmental requirements for the quiescence of adult somatic stem cells during their development from actively proliferating stem/progenitor cells.
MeSH term(s)	Animals ; Animals, Newborn ; Cell Culture Techniques ; Cell Differentiation ; Cell Hypoxia ; Cell Proliferation/drug effects ; Cells, Cultured ; Core Binding Factor Alpha 2 Subunit/metabolism ; Cyclin-Dependent Kinase Inhibitor p15/metabolism ; Kruppel-Like Transcription Factors/metabolism ; Mice ; Oligodendrocyte Precursor Cells/cytology ; Oligodendrocyte Precursor Cells/drug effects ; Oligodendrocyte Precursor Cells/metabolism ; Rats ; Signal Transduction ; Thyroid Hormones/pharmacology
Chemical Substances	Core Binding Factor Alpha 2 Subunit ; Cyclin-Dependent Kinase Inhibitor p15 ; Kruppel-Like Transcription Factors ; Thyroid Hormones
Language	English
Publishing date	2017-04-21
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-017-01023-9
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Article ; Online: CO-CBS-H2 S Axis: From Vascular Mediator to Cancer Regulator.

Suematsu, Makoto / Nakamura, Takashi / Tokumoto, Yasuhito / Yamamoto, Takehiro / Kajimura, Mayumi / Kabe, Yasuaki

Microcirculation (New York, N.Y. : 1994)

2016 Volume 23, Issue 3, Page(s) 183–190

Abstract: CO is a gaseous mediator generated by HO. Our previous studies revealed that CO generated from inducible HO-1 or from constitutive HO-2 modulates function of different heme proteins or enzymes through binding to their prosthetic ferrous heme to alter ... ...

Abstract	CO is a gaseous mediator generated by HO. Our previous studies revealed that CO generated from inducible HO-1 or from constitutive HO-2 modulates function of different heme proteins or enzymes through binding to their prosthetic ferrous heme to alter their structures, regulating biological function of cells and organs. Such CO-directed target macromolecules include sGC and CBS. In the liver, CO serves as a sinusoidal dilator through its action on sGC in hepatic stellate cells, while the same gas accounts for vasoconstrictor that inhibits H2S generated by CO-sensitive CBS in astrocytes. Since molecular O2 is a substrate for HO, the latter mechanism contributes to hypoxic vasodilation in neurovascular units. We have recently uncovered that stress-inducible CO in and around cancer cells suppresses CBS to result in decreased methylation of PFKFB3, the enzyme regulating PFK-1, leading to a shift of glucose biotransformation from glycolysis toward pentose phosphate pathway; such a metabolic remodeling causes chemoresistance through increasing NADPH and reduced glutathione under stress conditions for cancer cells. This article reviews the intriguing networks of CO-sensitive metabolic regulatory mechanisms in microcirculation and cancer.
MeSH term(s)	Animals ; Capillaries/metabolism ; Capillaries/pathology ; Carbon Monoxide/metabolism ; Cystathionine beta-Synthase/metabolism ; Heme Oxygenase (Decyclizing)/metabolism ; Heme Oxygenase-1/metabolism ; Humans ; Hydrogen Sulfide/metabolism ; Kupffer Cells/metabolism ; Kupffer Cells/pathology ; Liver/metabolism ; Liver/pathology ; Liver Neoplasms/metabolism ; Liver Neoplasms/pathology ; Neoplasm Proteins/metabolism ; Synaptic Transmission
Chemical Substances	Neoplasm Proteins ; Carbon Monoxide (7U1EE4V452) ; HMOX1 protein, human (EC 1.14.14.18) ; Heme Oxygenase (Decyclizing) (EC 1.14.14.18) ; Heme Oxygenase-1 (EC 1.14.14.18) ; heme oxygenase-2 (EC 1.14.14.18) ; Cystathionine beta-Synthase (EC 4.2.1.22) ; Hydrogen Sulfide (YY9FVM7NSN)
Language	English
Publishing date	2016-04
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	1217758-1
ISSN	1549-8719 ; 1073-9688
ISSN (online)	1549-8719
ISSN	1073-9688
DOI	10.1111/micc.12253
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Article: Comparison of efficiency of terminal differentiation of oligodendrocytes from induced pluripotent stem cells versus embryonic stem cells in vitro

Tokumoto, Yasuhito / Ogawa, Shinichiro / Nagamune, Teruyuki / Miyake, Jun

Journal of bioscience and bioengineering. 2010 June, v. 109, no. 6

2010

Abstract: Oligodendrocytes are the myelinating cells of the central nervous system (CNS), and defects in these cells can result in the loss of CNS functions. Although oligodendrocyte progenitor cells transplantation therapy is an effective cure for such symptoms, ... ...

Abstract	Oligodendrocytes are the myelinating cells of the central nervous system (CNS), and defects in these cells can result in the loss of CNS functions. Although oligodendrocyte progenitor cells transplantation therapy is an effective cure for such symptoms, there is no readily available source of these cells. Recent studies have described the generation of induced pluripotent stem cells (iPS cells) from somatic cells, leading to anticipation of this technique as a novel therapeutic tool in regenerative medicine. In this study, we evaluated the ability of iPS cells derived from mouse embryonic fibroblasts to differentiate into oligodendrocytes and compared this with the differential ability of mouse embryonic stem cells (ES cells). Experiments using an in vitro oligodendrocyte differentiation protocol that was optimized to ES cells demonstrated that 2.3% of iPS cells differentiated into O4⁺ oligodendrocytes compared with 24.0% of ES cells. However, the rate of induction of A2B5⁺ oligodendrocyte precursor cell (OPC) was similar for both iPS-derived cells and ES-derived cells (14.1% and 12.6%, respectively). These findings suggest that some intracellular factors in iPS cells inhibit the terminal differentiation of oligodendrocytes from the OPC stage.
Language	English
Dates of publication	2010-06
Size	p. 622-628.
Publishing place	Osaka, Japan: Society for Bioscience and Bioengineering, Japan; Amsterdam, the Netherlands: Distributed outside Japan by Elsevier Science
Document type	Article
ZDB-ID	1465387-4
ISSN	1347-4421 ; 1389-1723
ISSN (online)	1347-4421
ISSN	1389-1723
DOI	10.1016/j.jbiosc.2009.11.013
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Article ; Online: Parameter optimization by using differential elimination

Nakatsui Masahiko / Horimoto Katsuhisa / Okamoto Masahiro / Tokumoto Yasuhito / Miyake Jun

BMC Systems Biology, Vol 4, Iss Suppl 2, p S

a general approach for introducing constraints into objective functions

2010 Volume 9

Abstract: Abstract Background The investigation of network dynamics is a major issue in systems and synthetic biology. One of the essential steps in a dynamics investigation is the parameter estimation in the model that expresses biological phenomena. Indeed, ... ...

Abstract	Abstract Background The investigation of network dynamics is a major issue in systems and synthetic biology. One of the essential steps in a dynamics investigation is the parameter estimation in the model that expresses biological phenomena. Indeed, various techniques for parameter optimization have been devised and implemented in both free and commercial software. While the computational time for parameter estimation has been greatly reduced, due to improvements in calculation algorithms and the advent of high performance computers, the accuracy of parameter estimation has not been addressed. Results We propose a new approach for parameter optimization by using differential elimination, to estimate kinetic parameter values with a high degree of accuracy. First, we utilize differential elimination, which is an algebraic approach for rewriting a system of differential equations into another equivalent system, to derive the constraints between kinetic parameters from differential equations. Second, we estimate the kinetic parameters introducing these constraints into an objective function, in addition to the error function of the square difference between the measured and estimated data, in the standard parameter optimization method. To evaluate the ability of our method, we performed a simulation study by using the objective function with and without the newly developed constraints: the parameters in two models of linear and non-linear equations, under the assumption that only one molecule in each model can be measured, were estimated by using a genetic algorithm (GA) and particle swarm optimization (PSO). As a result, the introduction of new constraints was dramatically effective: the GA and PSO with new constraints could successfully estimate the kinetic parameters in the simulated models, with a high degree of accuracy, while the conventional GA and PSO methods without them frequently failed. Conclusions The introduction of new constraints in an objective function by using differential elimination resulted in the drastic improvement of the estimation accuracy in parameter optimization methods. The performance of our approach was illustrated by simulations of the parameter optimization for two models of linear and non-linear equations, which included unmeasured molecules, by two types of optimization techniques. As a result, our method is a promising development in parameter optimization.
Keywords	Biology (General) ; QH301-705.5
Subject code	620
Language	English
Publishing date	2010-09-01T00:00:00Z
Publisher	BioMed Central
Document type	Article ; Online
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Article ; Online: Comparison of efficiency of terminal differentiation of oligodendrocytes from induced pluripotent stem cells versus embryonic stem cells in vitro.

Tokumoto, Yasuhito / Ogawa, Shinichiro / Nagamune, Teruyuki / Miyake, Jun

Journal of bioscience and bioengineering

2010 Volume 109, Issue 6, Page(s) 622–628

Abstract	Oligodendrocytes are the myelinating cells of the central nervous system (CNS), and defects in these cells can result in the loss of CNS functions. Although oligodendrocyte progenitor cells transplantation therapy is an effective cure for such symptoms, there is no readily available source of these cells. Recent studies have described the generation of induced pluripotent stem cells (iPS cells) from somatic cells, leading to anticipation of this technique as a novel therapeutic tool in regenerative medicine. In this study, we evaluated the ability of iPS cells derived from mouse embryonic fibroblasts to differentiate into oligodendrocytes and compared this with the differential ability of mouse embryonic stem cells (ES cells). Experiments using an in vitro oligodendrocyte differentiation protocol that was optimized to ES cells demonstrated that 2.3% of iPS cells differentiated into O4(+) oligodendrocytes compared with 24.0% of ES cells. However, the rate of induction of A2B5(+) oligodendrocyte precursor cell (OPC) was similar for both iPS-derived cells and ES-derived cells (14.1% and 12.6%, respectively). These findings suggest that some intracellular factors in iPS cells inhibit the terminal differentiation of oligodendrocytes from the OPC stage.
MeSH term(s)	Animals ; Cell Differentiation ; Embryonic Stem Cells/cytology ; Embryonic Stem Cells/metabolism ; Fibroblasts/cytology ; Induced Pluripotent Stem Cells/cytology ; Induced Pluripotent Stem Cells/metabolism ; Intermediate Filament Proteins/metabolism ; Mice ; Nerve Tissue Proteins/metabolism ; Nestin ; Oligodendroglia/cytology ; Oligodendroglia/metabolism ; Transcription Factors/metabolism
Chemical Substances	Intermediate Filament Proteins ; Nerve Tissue Proteins ; Nes protein, mouse ; Nestin ; Transcription Factors
Language	English
Publishing date	2010-06
Publishing country	Japan
Document type	Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1465387-4
ISSN	1347-4421 ; 1389-1723
ISSN (online)	1347-4421
ISSN	1389-1723
DOI	10.1016/j.jbiosc.2009.11.013
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Article ; Online: TRAIL inhibited the cyclic AMP responsible element mediated gene expression.

Tokumoto, Yasuhito / Horimoto, Katsuhisa / Miyake, Jun

Biochemical and biophysical research communications

2009 Volume 381, Issue 4, Page(s) 533–536

Abstract: Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) not only causes apoptotic cell death in tumor cells, but also activates some transcription factors and affects several other cellular functions. In this study, we observed the effect ... ...

Abstract	Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) not only causes apoptotic cell death in tumor cells, but also activates some transcription factors and affects several other cellular functions. In this study, we observed the effect of administration of TRAIL on gene expression downstream of the cyclic AMP responsive element (CRE) enhancer by using the signal transduction reporter cis-element plasmid pCRE-d2EGFP. Western blotting showed that after administration of TRAIL, the expression level of reporter protein d2EGFP was down-regulated in NIH3T3 cells. To confirm the TRAIL-induced down-regulation of CRE enhancer controlled gene expression, DNA Chip time series analysis of the intrinsic genes expressed in NIH3T3 cells was carried out. As a result, the expression levels of six genes, which have CRE sequence in their promoter region, were slightly down-regulated within three hours after administration of TRAIL.
MeSH term(s)	Animals ; Cyclic AMP Response Element-Binding Protein/metabolism ; Down-Regulation ; Gene Expression/drug effects ; Mice ; NIH 3T3 Cells ; Oligonucleotide Array Sequence Analysis ; Response Elements/drug effects ; TNF-Related Apoptosis-Inducing Ligand/pharmacology
Chemical Substances	Cyclic AMP Response Element-Binding Protein ; TNF-Related Apoptosis-Inducing Ligand
Language	English
Publishing date	2009-04-17
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	205723-2
ISSN	1090-2104 ; 0006-291X ; 0006-291X
ISSN (online)	1090-2104 ; 0006-291X
ISSN	0006-291X
DOI	10.1016/j.bbrc.2009.02.076
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Article ; Online: Induction of oligodendrocyte differentiation from adult human fibroblast-derived induced pluripotent stem cells.

Ogawa, Shin-ichiro / Tokumoto, Yasuhito / Miyake, Jun / Nagamune, Teruyuki

In vitro cellular & developmental biology. Animal

2011 Volume 47, Issue 7, Page(s) 464–469

Abstract: Induced pluripotent stem cells (iPSCs) prepared from somatic cells might become a novel therapeutic tool in regenerative medicine, especially for the central nervous system (CNS). In this study, we attempted to induce O4-positive (O4(+)) oligodendrocytes ...

Abstract	Induced pluripotent stem cells (iPSCs) prepared from somatic cells might become a novel therapeutic tool in regenerative medicine, especially for the central nervous system (CNS). In this study, we attempted to induce O4-positive (O4(+)) oligodendrocytes from adult human fibroblast-derived iPSCs in vitro. We used two adult human iPSC cell lines, 201B7 and 253G1. 201B7 was induced by four-gene transduction (oct4, sox2, klf4, c-myc), and 253G1 was induced by three-gene transduction (oct4, sox2, klf4). We treated these cells with two in vitro oligodendrocyte-directed differentiation protocols that were optimized for human embryonic stem cells. One protocol used platelet-derived growth factor as the major mitogen for oligodendrocyte lineage cells, and the other protocol used epidermal growth factor (EGF) as the mitogen. Although the differentiation efficiency was low (less than 0.01%), we could induce O4(+) oligodendrocytes from 253G1 cells using the EGF-dependent differentiation protocol. This is the first report of the in vitro induction of oligodendrocytes differentiation from human iPSCs.
MeSH term(s)	Adult ; Animals ; Cell Culture Techniques ; Cell Differentiation/physiology ; Cell Line ; Female ; Fibroblasts/cytology ; Fibroblasts/physiology ; Humans ; Induced Pluripotent Stem Cells/cytology ; Induced Pluripotent Stem Cells/physiology ; Mice ; Oligodendroglia/cytology ; Oligodendroglia/physiology
Language	English
Publishing date	2011-06-22
Publishing country	Germany
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1077810-x
ISSN	1543-706X ; 0883-8364 ; 1071-2690
ISSN (online)	1543-706X
ISSN	0883-8364 ; 1071-2690
DOI	10.1007/s11626-011-9435-2
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