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{"title"=>"TGF-beta1 Does Not Induce Senescence of Multipotent Mesenchymal Stromal Cells and Has Similar Effects in Early and Late Passages", "type"=>"journal", "authors"=>[{"first_name"=>"Gudrun", "last_name"=>"Walenda", "scopus_author_id"=>"54901991800"}, {"first_name"=>"Khalid", "last_name"=>"Abnaof", "scopus_author_id"=>"55316939200"}, {"first_name"=>"Sylvia", "last_name"=>"Joussen", "scopus_author_id"=>"6507518223"}, {"first_name"=>"Steffen", "last_name"=>"Meurer", "scopus_author_id"=>"8701222000"}, {"first_name"=>"Hubert", "last_name"=>"Smeets", "scopus_author_id"=>"55402551800"}, {"first_name"=>"Björn", "last_name"=>"Rath", "scopus_author_id"=>"23111862600"}, {"first_name"=>"Kurt", "last_name"=>"Hoffmann", "scopus_author_id"=>"8545541400"}, {"first_name"=>"Holger", "last_name"=>"Fröhlich", "scopus_author_id"=>"7005180387"}, {"first_name"=>"Martin", "last_name"=>"Zenke", "scopus_author_id"=>"7005565721"}, {"first_name"=>"Ralf", "last_name"=>"Weiskirchen", "scopus_author_id"=>"7004155039"}, {"first_name"=>"Wolfgang", "last_name"=>"Wagner", "scopus_author_id"=>"35228094600"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84885795846", "doi"=>"10.1371/journal.pone.0077656", "pui"=>"370046588", "pmid"=>"24147049", "scopus"=>"2-s2.0-84885795846", "issn"=>"19326203"}, "id"=>"b2d2bd1f-1d38-324f-b9ec-837aa5bb97de", "abstract"=>"Transforming growth factor-beta 1 (TGF-β1) stimulates a broad range of effects which are cell type dependent, and it has been suggested to induce cellular senescence. On the other hand, long-term culture of multipotent mesenchymal stromal cells (MSCs) has a major impact on their cellular physiology and therefore it is well conceivable that the molecular events triggered by TGF-β1 differ considerably in cells of early and late passages. In this study, we analyzed the effect of TGF-β1 on and during replicative senescence of MSCs. Stimulation with TGF-β1 enhanced proliferation, induced a network like growth pattern and impaired adipogenic and osteogenic differentiation. TGF-β1 did not induce premature senescence. However, due to increased proliferation rates the cells reached replicative senescence earlier than untreated controls. This was also evident, when we analyzed senescence-associated DNA-methylation changes. Gene expression profiles of MSCs differed considerably at relatively early (P 3-5) and later passages (P 10). Nonetheless, relative gene expression differences provoked by TGF-β1 at individual time points or in a time course dependent manner (stimulation for 0, 1, 4 and 12 h) were very similar in MSCs of early and late passage. These results support the notion that TGF-β1 has major impact on MSC function, but it does not induce senescence and has similar molecular effects during culture expansion.", "link"=>"http://www.mendeley.com/research/tgfbeta1-not-induce-senescence-multipotent-mesenchymal-stromal-cells-similar-effects-early-late-pass-2", "reader_count"=>30, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Student > Doctoral Student"=>3, "Researcher"=>5, "Student > Ph. D. Student"=>13, "Student > Postgraduate"=>2, "Other"=>1, "Student > Master"=>2, "Student > Bachelor"=>1, "Professor"=>2}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Student > Doctoral Student"=>3, "Researcher"=>5, "Student > Ph. D. Student"=>13, "Student > Postgraduate"=>2, "Other"=>1, "Student > Master"=>2, "Student > Bachelor"=>1, "Professor"=>2}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>5, "Mathematics"=>1, "Medicine and Dentistry"=>5, "Agricultural and Biological Sciences"=>14, "Chemical Engineering"=>1, "Psychology"=>1, "Computer Science"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>5}, "Psychology"=>{"Psychology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>14}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}, "Mathematics"=>{"Mathematics"=>1}, "Unspecified"=>{"Unspecified"=>1}, "Chemical Engineering"=>{"Chemical Engineering"=>1}}, "reader_count_by_country"=>{"Canada"=>1, "United States"=>1, "Japan"=>1, "United Kingdom"=>1, "Germany"=>1, "Spain"=>1}, "group_count"=>2}

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• http://doi.org/10.1016/j.cytogfr.2018.06.002
• http://doi.org/10.3390/ijms20205002
• http://doi.org/10.1016/j.procbio.2016.04.017
• http://doi.org/10.1016/j.yexcr.2017.03.056
• http://doi.org/10.1186/s13287-020-1552-y
• http://doi.org/10.1517/14712598.2015.1009886
• http://doi.org/10.3390/metabo10020059
• http://doi.org/10.1016/j.bbrc.2014.07.045
• http://doi.org/10.1016/j.jcyt.2013.11.004
• http://doi.org/10.1016/j.ejcb.2016.11.003
• http://doi.org/10.1177/2041731419842431
• http://doi.org/10.1038/srep08973
• http://doi.org/10.1016/j.jtbi.2017.11.024
• http://doi.org/10.1073/pnas.1407097111
• http://doi.org/10.1101/cshperspect.a022145
• http://doi.org/10.1007/s11033-018-4522-z

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Figshare

• {"files"=>["https://ndownloader.figshare.com/files/1246428"], "description"=>"<p>MSCs were stimulated for 7 days with increasing concentrations of TGF-β1 and proliferation was estimated using the MTT assay (<b>A</b>; n = 5). MSCs were cultured with or without 1 ng/mL TGF-β1 until they stopped proliferation. Cumulative population doublings (cPDs) were calculated throughout culture expansion and depicted by symbols for each passage (<b>B</b>; n = 6). The average culture period until proliferation arrest is shorter if cells are continuously cultured with TGF-β1 (<b>C</b>; n = 6). Comparison of cPDs for the first seven passages for TGF-β1 treated and un-treated MSC revealed a proliferative advantage particularly in the initial three passages (<b>D</b>; n = 6). The maximal cPDs at the time of ultimate proliferation arrest were similar with and without TGF-β1 treatment (<b>E</b>; n = 6). SA-β-gal activity was measured in MSCs cultured either with or without TGF-β1 for 7 passages by histochemical analysis with X-gal staining (<b>E</b>) or flowcytometric analysis of C₁₂FDG (<b>F</b>; n = 3; *p < 0.05; **p < 0.01).</p>", "links"=>[], "tags"=>["short-"], "article_id"=>826696, "categories"=>["Biological Sciences"], "users"=>["Gudrun Walenda", "Khalid Abnaof", "Sylvia Joussen", "Steffen Meurer", "Hubert Smeets", "Björn Rath", "Kurt Hoffmann", "Holger Fröhlich", "Martin Zenke", "Ralf Weiskirchen", "Wolfgang Wagner"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0077656.g002", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Influence_of_TGF_946_1_on_short_and_long_term_expansion_and_senescence_/826696", "title"=>"Influence of TGF-β1 on short- and long-term expansion and senescence.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-17 04:04:42"}
• {"files"=>["https://ndownloader.figshare.com/files/1246432", "https://ndownloader.figshare.com/files/1246433"], "description"=>"<div><p>Transforming growth factor-beta 1 (TGF-β1) stimulates a broad range of effects which are cell type dependent, and it has been suggested to induce cellular senescence. On the other hand, long-term culture of multipotent mesenchymal stromal cells (MSCs) has a major impact on their cellular physiology and therefore it is well conceivable that the molecular events triggered by TGF-β1 differ considerably in cells of early and late passages. In this study, we analyzed the effect of TGF-β1 on and during replicative senescence of MSCs. Stimulation with TGF-β1 enhanced proliferation, induced a network like growth pattern and impaired adipogenic and osteogenic differentiation. TGF-β1 did not induce premature senescence. However, due to increased proliferation rates the cells reached replicative senescence earlier than untreated controls. This was also evident, when we analyzed senescence-associated DNA-methylation changes. Gene expression profiles of MSCs differed considerably at relatively early (P 3 - 5) and later passages (P 10). Nonetheless, relative gene expression differences provoked by TGF-β1 at individual time points or in a time course dependent manner (stimulation for 0, 1, 4 and 12 h) were very similar in MSCs of early and late passage. These results support the notion that TGF-β1 has major impact on MSC function, but it does not induce senescence and has similar molecular effects during culture expansion.</p> </div>", "links"=>[], "tags"=>["induce", "senescence", "multipotent", "mesenchymal", "stromal", "cells"], "article_id"=>826699, "categories"=>["Biological Sciences"], "users"=>["Gudrun Walenda", "Khalid Abnaof", "Sylvia Joussen", "Steffen Meurer", "Hubert Smeets", "Björn Rath", "Kurt Hoffmann", "Holger Fröhlich", "Martin Zenke", "Ralf Weiskirchen", "Wolfgang Wagner"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0077656.s001", "https://dx.doi.org/10.1371/journal.pone.0077656.s002"], "stats"=>{"downloads"=>11, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_TGF_beta1_Does_Not_Induce_Senescence_of_Multipotent_Mesenchymal_Stromal_Cells_and_Has_Similar_Effects_in_Early_and_Late_Passages_/826699", "title"=>"TGF-beta1 Does Not Induce Senescence of Multipotent Mesenchymal Stromal Cells and Has Similar Effects in Early and Late Passages", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-10-17 04:04:42"}
• {"files"=>["https://ndownloader.figshare.com/files/1246429"], "description"=>"<p>MSCs were cultured for three passages either with or without TGF-β1. Then the state of cellular senescence was tracked using our recently described Epigenetic-Senescence-Signature which is based on DNA-methylation (DNAm) changes at six specific CpG sites in the genome [<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0077656#B35\" target=\"_blank\">35</a>,<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0077656#B36\" target=\"_blank\">36</a>]. Predicted passage numbers and real passage numbers correlated well for controls, whereas the passage number was significantly overestimated for TGF-β1 treated cells (<b>A</b>; *p < 0.05). Comparison of predicted and real cumulative population doublings (cPDs) reflected the proliferative advantage with TGF-β1. Overall, the number of cPDs was slightly overestimated by the Epigenetic-Aging-Signature but this cannot be attributed to TGF-β1 stimulation (<b>B</b>). Symbols represent different MSC preparations that were cultured with (black color) or without TGF-β1 (grey color).</p>", "links"=>[], "tags"=>["senescence-associated", "dna-methylation"], "article_id"=>826697, "categories"=>["Biological Sciences"], "users"=>["Gudrun Walenda", "Khalid Abnaof", "Sylvia Joussen", "Steffen Meurer", "Hubert Smeets", "Björn Rath", "Kurt Hoffmann", "Holger Fröhlich", "Martin Zenke", "Ralf Weiskirchen", "Wolfgang Wagner"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0077656.g003", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_TGF_946_1_on_senescence_associated_DNA_methylation_signature_/826697", "title"=>"Effect of TGF-β1 on senescence-associated DNA-methylation signature.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-17 04:04:42"}
• {"files"=>["https://ndownloader.figshare.com/files/1246430"], "description"=>"<p>Hierarchical clustering of global gene expression profiles (Euclidean distance) revealed inter-donor variation, a close relationship of early and late passages, and continuous changes with TGF-β1 stimulation (<b>A</b>). This was also reflected by principal component analysis (<b>B</b>; components 1 [PC1] and 2 [PC2] are depicted). TGF-β1-induced gene expression changes were compared in MSCs of early passage (P3 - P5) and later passage (P10) upon stimulation for either 1, 4, or 12 hours. Some genes are predominantly induced in early passage (depicted in blue) or in late passage (depicted in red) but the induced gene expression changes were overall very similar in MSCs of early and later passage (<b>C</b>). </p>", "links"=>[], "tags"=>[], "article_id"=>826698, "categories"=>["Biological Sciences"], "users"=>["Gudrun Walenda", "Khalid Abnaof", "Sylvia Joussen", "Steffen Meurer", "Hubert Smeets", "Björn Rath", "Kurt Hoffmann", "Holger Fröhlich", "Martin Zenke", "Ralf Weiskirchen", "Wolfgang Wagner"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0077656.g004", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Gene_expression_changes_upon_TGF_946_1_treatment_in_early_and_later_passages_/826698", "title"=>"Gene expression changes upon TGF-β1 treatment in early and later passages.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-17 04:04:42"}
• {"files"=>["https://ndownloader.figshare.com/files/1246427"], "description"=>"<p>Treatment with 1 ng/mL TGF-β1 induces a network-like growth pattern of MSCs within 7 days, which is reversed if the cells are re-seeded in media without TGF-β1 (<b>A</b>; crystal violet staining of fixed cells). Immunophenotypic analysis of MSCs upon continuous culture either with or without TGF-β1 for 4 to 5 passages was performed by flow cytometry. Exemplary histograms are depicted and analysis of mean fluorescence intensity (normalized to auto-fluorescence) did not reveal significant differences upon treatment with TGF-β1 (<b>B</b>; n = 5). MSCs that had been cultured with or without 1 ng/mL TGF-β1 for 1 to 4 passages were differentiated towards chondrogenic, osteogenic and adipogenic lineage. Particularly adipogenic differentiation was impaired by TGF−β1 (<b>C</b>; n = 3). </p>", "links"=>[], "tags"=>["msc"], "article_id"=>826695, "categories"=>["Biological Sciences"], "users"=>["Gudrun Walenda", "Khalid Abnaof", "Sylvia Joussen", "Steffen Meurer", "Hubert Smeets", "Björn Rath", "Kurt Hoffmann", "Holger Fröhlich", "Martin Zenke", "Ralf Weiskirchen", "Wolfgang Wagner"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0077656.g001", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Influence_of_TGF_1_on_MSC_growth_and_in_vitro_differentiation_/826695", "title"=>"Influence of TGF-β1 on MSC growth and <i>in</i><i>vitro</i> differentiation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-17 04:04:42"}

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Relative Metric

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F1000Prime