In Vitro Analysis of Breast Cancer Cell Line Tumourspheres and Primary Human Breast Epithelia Mammospheres Demonstrates Inter- and Intrasphere Heterogeneity
Events
Loading … Spinner

Mendeley | Further Information

{"title"=>"In Vitro Analysis of Breast Cancer Cell Line Tumourspheres and Primary Human Breast Epithelia Mammospheres Demonstrates Inter- and Intrasphere Heterogeneity", "type"=>"journal", "authors"=>[{"first_name"=>"Chanel E.", "last_name"=>"Smart", "scopus_author_id"=>"7101677728"}, {"first_name"=>"Brian J.", "last_name"=>"Morrison", "scopus_author_id"=>"7101794431"}, {"first_name"=>"Jodi M.", "last_name"=>"Saunus", "scopus_author_id"=>"24830736400"}, {"first_name"=>"Ana Cristina", "last_name"=>"Vargas", "scopus_author_id"=>"26640855900"}, {"first_name"=>"Patricia", "last_name"=>"Keith", "scopus_author_id"=>"8613616000"}, {"first_name"=>"Lynne", "last_name"=>"Reid", "scopus_author_id"=>"7201751485"}, {"first_name"=>"Leesa", "last_name"=>"Wockner", "scopus_author_id"=>"16306031000"}, {"first_name"=>"Marjan Askarian", "last_name"=>"Amiri", "scopus_author_id"=>"55994811900"}, {"first_name"=>"Debina", "last_name"=>"Sarkar", "scopus_author_id"=>"55757023700"}, {"first_name"=>"Peter T.", "last_name"=>"Simpson", "scopus_author_id"=>"7201907963"}, {"first_name"=>"Catherine", "last_name"=>"Clarke", "scopus_author_id"=>"7401438407"}, {"first_name"=>"Chris W.", "last_name"=>"Schmidt", "scopus_author_id"=>"24473520700"}, {"first_name"=>"Brent A.", "last_name"=>"Reynolds", "scopus_author_id"=>"7102514108"}, {"first_name"=>"Sunil R.", "last_name"=>"Lakhani", "scopus_author_id"=>"7004833949"}, {"first_name"=>"J. Alejandro", "last_name"=>"Lopez", "scopus_author_id"=>"13003441700"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"23750209", "scopus"=>"2-s2.0-84878623584", "doi"=>"10.1371/journal.pone.0064388", "sgr"=>"84878623584", "pui"=>"369060515"}, "id"=>"0a1e568c-0792-3096-9cda-cb97dbe60db6", "abstract"=>"Mammosphere and breast tumoursphere culture have gained popularity as in vitro assays for propagating and analysing normal and cancer stem cells. Whether the spheres derived from different sources or parent cultures themselves are indeed single entities enriched in stem/progenitor cells compared to other culture formats has not been fully determined. We surveyed sphere-forming capacity across 26 breast cell lines, immunophenotyped spheres from six luminal- and basal-like lines by immunohistochemistry and flow cytometry and compared clonogenicity between sphere, adherent and matrigel culture formats using in vitro functional assays. Analyses revealed morphological and molecular intra- and inter-sphere heterogeneity, consistent with adherent parental cell line phenotypes. Flow cytometry showed sphere culture does not universally enrich for markers previously associated with stem cell phenotypes, although we found some cell-line specific changes between sphere and adherent formats. Sphere-forming efficiency was significantly lower than adherent or matrigel clonogenicity and constant over serial passage. Surprisingly, self-renewal capacity of sphere-derived cells was similar/lower than other culture formats. We observed significant correlation between long-term-proliferating-cell symmetric division rates in sphere and adherent cultures, suggesting functional overlap between the compartments sustaining them. Experiments with normal primary human mammary epithelia, including sorted luminal (MUC1(+)) and basal/myoepithelial (CD10(+)) cells revealed distinct luminal-like, basal-like and mesenchymal entities amongst primary mammospheres. Morphological and colony-forming-cell assay data suggested mammosphere culture may enrich for a luminal progenitor phenotype, or induce reversion/relaxation of the basal/mesenchymal in vitro selection occurring with adherent culture. Overall, cell line tumourspheres and primary mammospheres are not homogenous entities enriched for stem cells, suggesting a more cautious approach to interpreting data from these assays and careful consideration of its limitations. Sphere culture may represent an alternative 3-dimensional culture system which rather than universally 'enriching' for stem cells, has utility as one of a suite of functional assays that provide a read-out of progenitor activity.", "link"=>"http://www.mendeley.com/research/vitro-analysis-breast-cancer-cell-line-tumourspheres-primary-human-breast-epithelia-mammospheres-dem", "reader_count"=>96, "reader_count_by_academic_status"=>{"Unspecified"=>3, "Professor > Associate Professor"=>4, "Researcher"=>25, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>30, "Student > Postgraduate"=>5, "Other"=>7, "Student > Master"=>10, "Student > Bachelor"=>5, "Lecturer"=>2, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>3, "Professor > Associate Professor"=>4, "Researcher"=>25, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>30, "Student > Postgraduate"=>5, "Other"=>7, "Student > Master"=>10, "Student > Bachelor"=>5, "Lecturer"=>2, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>4, "Biochemistry, Genetics and Molecular Biology"=>12, "Agricultural and Biological Sciences"=>52, "Medicine and Dentistry"=>15, "Pharmacology, Toxicology and Pharmaceutical Science"=>3, "Physics and Astronomy"=>1, "Chemical Engineering"=>1, "Chemistry"=>4, "Psychology"=>1, "Computer Science"=>1, "Immunology and Microbiology"=>2}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>15}, "Chemistry"=>{"Chemistry"=>4}, "Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Psychology"=>{"Psychology"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>52}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>12}, "Unspecified"=>{"Unspecified"=>4}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>3}, "Chemical Engineering"=>{"Chemical Engineering"=>1}}, "reader_count_by_country"=>{"Netherlands"=>1, "Luxembourg"=>1, "Italy"=>1, "United Kingdom"=>2, "South Africa"=>1, "France"=>1, "Portugal"=>1, "Spain"=>1}, "group_count"=>2}

Scopus | Further Information

{"@_fa"=>"true", "link"=>[{"@_fa"=>"true", "@ref"=>"self", "@href"=>"https://api.elsevier.com/content/abstract/scopus_id/84878623584"}, {"@_fa"=>"true", "@ref"=>"author-affiliation", "@href"=>"https://api.elsevier.com/content/abstract/scopus_id/84878623584?field=author,affiliation"}, {"@_fa"=>"true", "@ref"=>"scopus", "@href"=>"https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84878623584&origin=inward"}, {"@_fa"=>"true", "@ref"=>"scopus-citedby", "@href"=>"https://www.scopus.com/inward/citedby.uri?partnerID=HzOxMe3b&scp=84878623584&origin=inward"}], "prism:url"=>"https://api.elsevier.com/content/abstract/scopus_id/84878623584", "dc:identifier"=>"SCOPUS_ID:84878623584", "eid"=>"2-s2.0-84878623584", "dc:title"=>"In Vitro Analysis of Breast Cancer Cell Line Tumourspheres and Primary Human Breast Epithelia Mammospheres Demonstrates Inter- and Intrasphere Heterogeneity", "dc:creator"=>"Smart C.", "prism:publicationName"=>"PLoS ONE", "prism:eIssn"=>"19326203", "prism:volume"=>"8", "prism:issueIdentifier"=>"6", "prism:pageRange"=>nil, "prism:coverDate"=>"2013-06-04", "prism:coverDisplayDate"=>"4 June 2013", "prism:doi"=>"10.1371/journal.pone.0064388", "citedby-count"=>"38", "affiliation"=>[{"@_fa"=>"true", "affilname"=>"UQ Centre for Clinical Research", "affiliation-city"=>"Brisbane", "affiliation-country"=>"Australia"}, {"@_fa"=>"true", "affilname"=>"QIMR Berghofer Medical Research Institute", "affiliation-city"=>"Brisbane", "affiliation-country"=>"Australia"}], "pubmed-id"=>"23750209", "prism:aggregationType"=>"Journal", "subtype"=>"ar", "subtypeDescription"=>"Article", "article-number"=>"e64388", "source-id"=>"10600153309", "openaccess"=>"1", "openaccessFlag"=>true}

Facebook

  • {"url"=>"http%3A%2F%2Fjournals.plos.org%2Fplosone%2Farticle%3Fid%3D10.1371%252Fjournal.pone.0064388", "share_count"=>0, "like_count"=>0, "comment_count"=>0, "click_count"=>0, "total_count"=>0}

Counter

  • {"month"=>"6", "year"=>"2013", "pdf_views"=>"154", "xml_views"=>"6", "html_views"=>"551"}
  • {"month"=>"7", "year"=>"2013", "pdf_views"=>"57", "xml_views"=>"2", "html_views"=>"187"}
  • {"month"=>"8", "year"=>"2013", "pdf_views"=>"69", "xml_views"=>"0", "html_views"=>"188"}
  • {"month"=>"9", "year"=>"2013", "pdf_views"=>"59", "xml_views"=>"0", "html_views"=>"183"}
  • {"month"=>"10", "year"=>"2013", "pdf_views"=>"48", "xml_views"=>"2", "html_views"=>"205"}
  • {"month"=>"11", "year"=>"2013", "pdf_views"=>"48", "xml_views"=>"1", "html_views"=>"140"}
  • {"month"=>"12", "year"=>"2013", "pdf_views"=>"44", "xml_views"=>"0", "html_views"=>"149"}
  • {"month"=>"1", "year"=>"2014", "pdf_views"=>"27", "xml_views"=>"0", "html_views"=>"89"}
  • {"month"=>"2", "year"=>"2014", "pdf_views"=>"44", "xml_views"=>"0", "html_views"=>"192"}
  • {"month"=>"3", "year"=>"2014", "pdf_views"=>"51", "xml_views"=>"2", "html_views"=>"228"}
  • {"month"=>"4", "year"=>"2014", "pdf_views"=>"35", "xml_views"=>"3", "html_views"=>"184"}
  • {"month"=>"5", "year"=>"2014", "pdf_views"=>"55", "xml_views"=>"0", "html_views"=>"177"}
  • {"month"=>"6", "year"=>"2014", "pdf_views"=>"38", "xml_views"=>"0", "html_views"=>"126"}
  • {"month"=>"7", "year"=>"2014", "pdf_views"=>"49", "xml_views"=>"1", "html_views"=>"146"}
  • {"month"=>"8", "year"=>"2014", "pdf_views"=>"26", "xml_views"=>"2", "html_views"=>"120"}
  • {"month"=>"9", "year"=>"2014", "pdf_views"=>"29", "xml_views"=>"1", "html_views"=>"146"}
  • {"month"=>"10", "year"=>"2014", "pdf_views"=>"40", "xml_views"=>"2", "html_views"=>"150"}
  • {"month"=>"11", "year"=>"2014", "pdf_views"=>"27", "xml_views"=>"1", "html_views"=>"114"}
  • {"month"=>"12", "year"=>"2014", "pdf_views"=>"23", "xml_views"=>"2", "html_views"=>"99"}
  • {"month"=>"1", "year"=>"2015", "pdf_views"=>"31", "xml_views"=>"1", "html_views"=>"179"}
  • {"month"=>"2", "year"=>"2015", "pdf_views"=>"27", "xml_views"=>"0", "html_views"=>"147"}
  • {"month"=>"3", "year"=>"2015", "pdf_views"=>"28", "xml_views"=>"0", "html_views"=>"148"}
  • {"month"=>"4", "year"=>"2015", "pdf_views"=>"21", "xml_views"=>"0", "html_views"=>"154"}
  • {"month"=>"5", "year"=>"2015", "pdf_views"=>"32", "xml_views"=>"1", "html_views"=>"212"}
  • {"month"=>"6", "year"=>"2015", "pdf_views"=>"35", "xml_views"=>"0", "html_views"=>"181"}
  • {"month"=>"7", "year"=>"2015", "pdf_views"=>"28", "xml_views"=>"0", "html_views"=>"209"}
  • {"month"=>"8", "year"=>"2015", "pdf_views"=>"24", "xml_views"=>"1", "html_views"=>"149"}
  • {"month"=>"9", "year"=>"2015", "pdf_views"=>"31", "xml_views"=>"0", "html_views"=>"165"}
  • {"month"=>"10", "year"=>"2015", "pdf_views"=>"34", "xml_views"=>"0", "html_views"=>"193"}
  • {"month"=>"11", "year"=>"2015", "pdf_views"=>"32", "xml_views"=>"0", "html_views"=>"197"}
  • {"month"=>"12", "year"=>"2015", "pdf_views"=>"40", "xml_views"=>"0", "html_views"=>"152"}
  • {"month"=>"1", "year"=>"2016", "pdf_views"=>"16", "xml_views"=>"0", "html_views"=>"120"}
  • {"month"=>"2", "year"=>"2016", "pdf_views"=>"6", "xml_views"=>"0", "html_views"=>"202"}
  • {"month"=>"3", "year"=>"2016", "pdf_views"=>"31", "xml_views"=>"0", "html_views"=>"160"}
  • {"month"=>"4", "year"=>"2016", "pdf_views"=>"45", "xml_views"=>"0", "html_views"=>"155"}
  • {"month"=>"5", "year"=>"2016", "pdf_views"=>"29", "xml_views"=>"0", "html_views"=>"116"}
  • {"month"=>"6", "year"=>"2016", "pdf_views"=>"42", "xml_views"=>"0", "html_views"=>"146"}
  • {"month"=>"7", "year"=>"2016", "pdf_views"=>"30", "xml_views"=>"0", "html_views"=>"156"}
  • {"month"=>"8", "year"=>"2016", "pdf_views"=>"29", "xml_views"=>"0", "html_views"=>"135"}
  • {"month"=>"9", "year"=>"2016", "pdf_views"=>"31", "xml_views"=>"0", "html_views"=>"161"}
  • {"month"=>"10", "year"=>"2016", "pdf_views"=>"31", "xml_views"=>"0", "html_views"=>"132"}
  • {"month"=>"11", "year"=>"2016", "pdf_views"=>"11", "xml_views"=>"0", "html_views"=>"154"}
  • {"month"=>"12", "year"=>"2016", "pdf_views"=>"19", "xml_views"=>"1", "html_views"=>"115"}
  • {"month"=>"1", "year"=>"2017", "pdf_views"=>"31", "xml_views"=>"0", "html_views"=>"162"}
  • {"month"=>"2", "year"=>"2017", "pdf_views"=>"30", "xml_views"=>"0", "html_views"=>"162"}
  • {"month"=>"3", "year"=>"2017", "pdf_views"=>"45", "xml_views"=>"3", "html_views"=>"187"}
  • {"month"=>"4", "year"=>"2017", "pdf_views"=>"33", "xml_views"=>"0", "html_views"=>"208"}
  • {"month"=>"5", "year"=>"2017", "pdf_views"=>"61", "xml_views"=>"1", "html_views"=>"204"}
  • {"month"=>"6", "year"=>"2017", "pdf_views"=>"42", "xml_views"=>"0", "html_views"=>"169"}
  • {"month"=>"7", "year"=>"2017", "pdf_views"=>"44", "xml_views"=>"3", "html_views"=>"147"}
  • {"month"=>"8", "year"=>"2017", "pdf_views"=>"31", "xml_views"=>"0", "html_views"=>"152"}
  • {"month"=>"9", "year"=>"2017", "pdf_views"=>"22", "xml_views"=>"2", "html_views"=>"142"}
  • {"month"=>"10", "year"=>"2017", "pdf_views"=>"28", "xml_views"=>"2", "html_views"=>"202"}
  • {"month"=>"11", "year"=>"2017", "pdf_views"=>"37", "xml_views"=>"1", "html_views"=>"189"}
  • {"month"=>"12", "year"=>"2017", "pdf_views"=>"15", "xml_views"=>"2", "html_views"=>"115"}
  • {"month"=>"1", "year"=>"2018", "pdf_views"=>"24", "xml_views"=>"1", "html_views"=>"121"}
  • {"month"=>"2", "year"=>"2018", "pdf_views"=>"20", "xml_views"=>"0", "html_views"=>"102"}
  • {"month"=>"3", "year"=>"2018", "pdf_views"=>"36", "xml_views"=>"2", "html_views"=>"126"}
  • {"month"=>"4", "year"=>"2018", "pdf_views"=>"21", "xml_views"=>"0", "html_views"=>"81"}
  • {"month"=>"5", "year"=>"2018", "pdf_views"=>"31", "xml_views"=>"0", "html_views"=>"92"}
  • {"month"=>"6", "year"=>"2018", "pdf_views"=>"24", "xml_views"=>"0", "html_views"=>"86"}
  • {"month"=>"7", "year"=>"2018", "pdf_views"=>"12", "xml_views"=>"5", "html_views"=>"61"}
  • {"month"=>"8", "year"=>"2018", "pdf_views"=>"27", "xml_views"=>"1", "html_views"=>"104"}
  • {"month"=>"9", "year"=>"2018", "pdf_views"=>"19", "xml_views"=>"0", "html_views"=>"90"}
  • {"month"=>"10", "year"=>"2018", "pdf_views"=>"28", "xml_views"=>"1", "html_views"=>"77"}
  • {"month"=>"11", "year"=>"2018", "pdf_views"=>"18", "xml_views"=>"1", "html_views"=>"76"}
  • {"month"=>"12", "year"=>"2018", "pdf_views"=>"8", "xml_views"=>"1", "html_views"=>"71"}
  • {"month"=>"1", "year"=>"2019", "pdf_views"=>"29", "xml_views"=>"0", "html_views"=>"55"}
  • {"month"=>"2", "year"=>"2019", "pdf_views"=>"22", "xml_views"=>"0", "html_views"=>"63"}
  • {"month"=>"3", "year"=>"2019", "pdf_views"=>"20", "xml_views"=>"1", "html_views"=>"61"}
  • {"month"=>"4", "year"=>"2019", "pdf_views"=>"17", "xml_views"=>"0", "html_views"=>"57"}
  • {"month"=>"5", "year"=>"2019", "pdf_views"=>"27", "xml_views"=>"0", "html_views"=>"69"}
  • {"month"=>"6", "year"=>"2019", "pdf_views"=>"33", "xml_views"=>"0", "html_views"=>"80"}
  • {"month"=>"7", "year"=>"2019", "pdf_views"=>"28", "xml_views"=>"0", "html_views"=>"65"}
  • {"month"=>"8", "year"=>"2019", "pdf_views"=>"34", "xml_views"=>"0", "html_views"=>"63"}
  • {"month"=>"9", "year"=>"2019", "pdf_views"=>"19", "xml_views"=>"0", "html_views"=>"53"}
  • {"month"=>"10", "year"=>"2019", "pdf_views"=>"20", "xml_views"=>"0", "html_views"=>"54"}
  • {"month"=>"11", "year"=>"2019", "pdf_views"=>"27", "xml_views"=>"0", "html_views"=>"41"}
  • {"month"=>"12", "year"=>"2019", "pdf_views"=>"25", "xml_views"=>"0", "html_views"=>"56"}
  • {"month"=>"1", "year"=>"2020", "pdf_views"=>"15", "xml_views"=>"0", "html_views"=>"46"}
  • {"month"=>"2", "year"=>"2020", "pdf_views"=>"22", "xml_views"=>"1", "html_views"=>"57"}
  • {"month"=>"3", "year"=>"2020", "pdf_views"=>"19", "xml_views"=>"0", "html_views"=>"42"}
  • {"month"=>"4", "year"=>"2020", "pdf_views"=>"45", "xml_views"=>"1", "html_views"=>"24"}
  • {"month"=>"5", "year"=>"2020", "pdf_views"=>"72", "xml_views"=>"1", "html_views"=>"26"}
  • {"month"=>"6", "year"=>"2020", "pdf_views"=>"15", "xml_views"=>"1", "html_views"=>"20"}
  • {"month"=>"7", "year"=>"2020", "pdf_views"=>"9", "xml_views"=>"0", "html_views"=>"28"}
  • {"month"=>"8", "year"=>"2020", "pdf_views"=>"24", "xml_views"=>"1", "html_views"=>"31"}
  • {"month"=>"9", "year"=>"2020", "pdf_views"=>"8", "xml_views"=>"2", "html_views"=>"35"}
  • {"month"=>"10", "year"=>"2020", "pdf_views"=>"20", "xml_views"=>"1", "html_views"=>"24"}
  • {"month"=>"11", "year"=>"2020", "pdf_views"=>"37", "xml_views"=>"0", "html_views"=>"24"}
  • {"month"=>"12", "year"=>"2020", "pdf_views"=>"4", "xml_views"=>"1", "html_views"=>"1"}

Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1074311"], "description"=>"<p>(A) Light microscope images of cell lines grown in adherent and sphere-promoting suspension conditions 7 d after seeding (images taken at 40× magnification, insets 100x magnification). H&E analysis of sections of formalin-fixed, paraffin embedded spheres are also shown. Arrows indicate wide lumina within MCF7 spheres and cleft-like lumina within KPL-1 spheres. Scale bars 100 µm. (B) Average sphere diameter measured by digital image measurement of the long axis of spheres. Data represent median diameter of spheres ± SD measured in 10 random fields, representative of two biological replicates. (C) Light microscope images of structures formed by MCF7 and SUM-159-PT cells grown in matrigel. Images taken at 100x magnification, scale bar 100 µm. (D) Representative images from immunohistochemical analysis of the indicated antigens on FFPE preparations of spheres from basal-like (i, iii, ix, xv, xvii: SUM-159-PT; v, vii, xi, xiii: HBL100) and luminal-like cell lines (ii, vi, xii, xiv: MCF7; iv, viii, xvi, xviii: KPL-1; x: BT-474). Complete data set available in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0064388#pone.0064388.s001\" target=\"_blank\">Figure S1</a>. Images taken at 200x magnification (black triangle, 400x). Scale bar 100 µm. Black arrows indicate intermittent laminin1/2.</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Adult stem cells", "Stem cell lines", "Cell differentiation", "morphogenesis", "histology", "Molecular cell biology", "Cellular types", "Epithelial cells", "Cell growth", "immunohistochemical", "tumourspheres", "cultured", "luminal-like", "cancer"], "article_id"=>711562, "categories"=>["Biological Sciences"], "users"=>["Chanel E. Smart", "Brian J. Morrison", "Jodi M. Saunus", "Ana Cristina Vargas", "Patricia Keith", "Lynne Reid", "Leesa Wockner", "Marjan Askarian Amiri", "Debina Sarkar", "Peter T. Simpson", "Catherine Clarke", "Chris W. Schmidt", "Brent A. Reynolds", "Sunil R. Lakhani", "J. Alejandro Lopez"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0064388.g001", "stats"=>{"downloads"=>1, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Morphologic_and_immunohistochemical_analysis_of_tumourspheres_cultured_from_a_panel_of_basal_like_claudin_low_and_luminal_like_breast_cancer_cell_lines_/711562", "title"=>"Morphologic and immunohistochemical analysis of tumourspheres cultured from a panel of basal-like/claudin-low and luminal-like breast cancer cell lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-04 00:26:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/1074316"], "description"=>"<p>(<b>A</b>) <b>Experiment design.</b> Matched adherent and sphere cultures of luminal-like and basal-like/claudin-low cell lines were generated in triplicate from subconfluent, adherent parent cultures on d0, harvested and dissociated on d7 into single cell suspensions, then concurrently stained with combinations of fluorescent antibody conjugates. Panel 1: MUC1-FITC, HER2-PE and the LIVE/DEAD® red cell viability stain; Panel 2: CD49f-Pacific Blue, Aldefluor® reagent (for ALDH1 activity), CD24-PE, LIVE/DEAD® red, AC133-APC, EpCAM-PerCP Cy5.5 and CD44-APC-Cy7. (<b>B</b>) <b>Gating strategy for simultaneous detection of cell surface markers.</b> Acellular particles and dead cells were excluded based on low light scatter and LIVE/DEAD® red positivity (live cells designated ‘LDR live’). For each experiment, gates were placed based on autofluorescence of unstained adherent or sphere control samples to account for differences in the autofluorescence of cells grown in the different formats. For Aldefluor® (ALDH1 activity), gates were placed based on the fluorescence of the DEAB negative control (refer to supp methods). Population frequencies were determined for individual parameters (MUC1+, HER2+, AC133+, EpCAM+, ALDH1+, CD44+, CD24+, CD49f+). For panel 2, combination gating was performed to investigate the frequencies of stem cell populations, differentiation states and other subpopulations (see Fig. 3).</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Adult stem cells", "Stem cell lines", "Cell differentiation", "morphogenesis", "histology", "Molecular cell biology", "Cellular types", "Epithelial cells", "Cell growth", "cytometry", "mammary", "epithelial", "markers", "matched", "adherent", "cultures", "cancer"], "article_id"=>711567, "categories"=>["Biological Sciences"], "users"=>["Chanel E. Smart", "Brian J. Morrison", "Jodi M. Saunus", "Ana Cristina Vargas", "Patricia Keith", "Lynne Reid", "Leesa Wockner", "Marjan Askarian Amiri", "Debina Sarkar", "Peter T. Simpson", "Catherine Clarke", "Chris W. Schmidt", "Brent A. Reynolds", "Sunil R. Lakhani", "J. Alejandro Lopez"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0064388.g002", "stats"=>{"downloads"=>5, "page_views"=>127, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Multiparametric_flow_cytometry_analysis_of_mammary_epithelial_and_stem_cell_markers_in_matched_adherent_and_sphere_cultures_of_breast_cancer_cell_lines_/711567", "title"=>"Multiparametric flow cytometry analysis of mammary epithelial and stem cell markers in matched adherent and sphere cultures of breast cancer cell lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-04 00:26:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/1074318"], "description"=>"<p>Adherent and sphere cultures were dissociated, stained with fluorescent antibody conjugates and analysed as described in Fig. 2. Heat maps show changes in subpopulation frequencies within the live cell population. Red shades = higher frequency in spheres compared to matched adherent cultures; blue shades = lower frequency in spheres; empty = not consistent and/or not statistically significant across biological replicates; diagonal line = not determined. Statistical significance was determined using two-way ANOVA and 2-tailed, paired t-tests. Results are depicted only where the trend was directionally consistent and statistically significant over biological replicates (refer to methods). Significance levels are colour-coded: light pink/blue: <i>P</i> = 0.05–0.001; mid-pink/blue: <i>P</i> = 0.001–0.0001; dark red/blue: <i>P</i><0.0001). Dotted boxes indicate changes of interest: significant enrichment with AC133+ cells in 3/6 cell lines (A), and frequent shifts in CD49f<sup>+</sup>/CD24<sup>+</sup> phenotypes with sphere culture (B).</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Adult stem cells", "Stem cell lines", "Cell differentiation", "morphogenesis", "histology", "Molecular cell biology", "Cellular types", "Epithelial cells", "Cell growth", "frequencies", "mammary", "epithelial", "subpopulations", "parallel", "adherent", "cultures", "cancer", "lines", "multiparametric", "cytometry"], "article_id"=>711569, "categories"=>["Biological Sciences"], "users"=>["Chanel E. Smart", "Brian J. Morrison", "Jodi M. Saunus", "Ana Cristina Vargas", "Patricia Keith", "Lynne Reid", "Leesa Wockner", "Marjan Askarian Amiri", "Debina Sarkar", "Peter T. Simpson", "Catherine Clarke", "Chris W. Schmidt", "Brent A. Reynolds", "Sunil R. Lakhani", "J. Alejandro Lopez"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0064388.g003", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Differences_in_the_frequencies_of_functional_mammary_epithelial_subpopulations_in_parallel_sphere_and_adherent_cultures_of_breast_cancer_cell_lines_assessed_by_multiparametric_flow_cytometry_analysis_/711569", "title"=>"Differences in the frequencies of functional mammary epithelial subpopulations in parallel sphere and adherent cultures of breast cancer cell lines assessed by multiparametric flow cytometry analysis.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-04 00:26:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/1074321"], "description"=>"<p><b>(A) Sphere forming efficiency (SFE) of cell lines through serial sphere passage.</b>–14 replicates per dilution, 4 dilutions/cell line/passage), expressed as a percentage of the original number of cells seeded, then averaged across the dilutions. Bars represent the mean SFE ± SD from 2–3 independent experiments. Two-tailed, unpaired t-tests demonstrated no significant difference in SFE between passage 1 and 5 for any cell line (<i>ns</i>). (<b>B</b>) <b>Correlation between stem cell symmetric division rates of matched adherent and sphere cultures.</b> Ten breast cancer cell lines (MCF7, KPL-1, BT-474, SKBR-3, T47D, SUM-159-PT, MDA-MB-436, HBL100, Hs578T and BT-20) were seeded in triplicate at equal densities in adherent or sphere-promoting conditions, and the total number of cells was calculated every 5–7 d for 2–8 passages. Mean fold expansion and the rates of long-term proliferating cell symmetric division (K<i><sub>ll</sub></i>) were calculated for each culture <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0064388#pone.0064388-Deleyrolle1\" target=\"_blank\">[30]</a> and plotted for correlation analysis (solid line, linear correlation; dotted lines 95% confidence interval). The relationship between adherent and sphere K<i><sub>II</sub></i> rates was statistically significant (linear correlation analysis (post-test for linear trend); <i>P</i> = 0.0016). (<b>C</b>) <b>Primary clonogenicity of MCF7 and SUM-159-PT cells in different growth formats.</b> Clonogenicity in adherent growth conditions, matrigel overlay and sphere-promoting conditions was calculated as the number of colonies (adherent or non-adherent) that grew after 7 d, as a percentage of cells seeded. Data shown are means ± SE, representative of at least three separate experiments performed in triplicate. (<b>D</b>) <b>Secondary clonogenicity of MCF7 and SUM-159-PT cells derived from spheres and matrigel structures.</b> Clonogenicity was calculated after dissociating and reseeding cells derived from spheres and matrigel structures into adherent conditions at limiting dilution. Data are means ± SE, representative of one of three separate experiments performed in triplicate. <i>P</i> values in C and D were generated using two-tailed, unpaired t-tests.</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Adult stem cells", "Stem cell lines", "Cell differentiation", "morphogenesis", "histology", "Molecular cell biology", "Cellular types", "Epithelial cells", "Cell growth", "progenitor", "cancer"], "article_id"=>711572, "categories"=>["Biological Sciences"], "users"=>["Chanel E. Smart", "Brian J. Morrison", "Jodi M. Saunus", "Ana Cristina Vargas", "Patricia Keith", "Lynne Reid", "Leesa Wockner", "Marjan Askarian Amiri", "Debina Sarkar", "Peter T. Simpson", "Catherine Clarke", "Chris W. Schmidt", "Brent A. Reynolds", "Sunil R. Lakhani", "J. Alejandro Lopez"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0064388.g004", "stats"=>{"downloads"=>2, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_vitro_functional_analysis_of_progenitor_cell_content_in_breast_cancer_cell_line_tumourspheres_/711572", "title"=>"<i>In vitro</i> functional analysis of progenitor cell content in breast cancer cell line tumourspheres.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-04 00:26:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/1074322"], "description"=>"<p>(<b>A</b>) <b>Strategy for isolating luminal and myoepithelial progenitor-enriched subpopulations from reduction mammoplasty tissue.</b> Tissues were physically and enzymatically processed to epithelial-rich, single cell suspensions, then cultured for 7d in mammary epithelial growth medium to generate enough cells for fluorescence-activated cell sorting (FACS) and sphere culture. Primary cultures were stained with fluorescent antibody conjugates (MUC1-FITC, CD10-PE-Cy5), the Sytox® Blue viability stain and ‘Lineage’ cocktail (‘Lin’: CD140b-PE, CD45-PE, CD31-PE; markers of stromal fibroblasts, leukocytes and endothelia respectively). CD10 and MUC1-sorted cells were placed in sphere-promoting culture for 10d before analysis. (<b>B</b>) <b>Gating strategy for enrichment of luminal- and myoepithelial-like progenitor cells.</b> Acellular particles and dead cells were excluded based on low light scatter and Sytox® blue positivity. Non-epithelial, PE<sup>+</sup> cells were also excluded, then MUC1<sup>+</sup> and CD10<sup>+</sup> cells were collected for sphere culture. (<b>C</b>) <b>Light microscope images (i,iii) and H&E histological sections (ii,iv–vii) of spheres generated from CD10<sup>+</sup> or MUC1<sup>+</sup> primary breast epithelial cells.</b> Open arrows, MUC1+-derived spheres often had limited lateral connections giving a petal-like appearance. Solid arrows, shows single cell with signet ring secretory morphology. (<b>D</b>) <b>Immunophenotypic analysis of MUC1<sup>+</sup> and CD10<sup>+</sup> progenitor-derived primary breast mammospheres.</b> Spheres were generated as described in A/B. Representative images from immunohistochemical analysis of the indicated antigens on FFPE preparations of spheres are shown. Images taken at 200× magnification. Scale bar 100 µm. Black arrows indicate apical membranous staining of E-cadherin and EGFR in luminal-like spheres. Red arrows indicate the mesenchymal-like spheres found rarely amongst the dominant structures formed in both CD10+ and MUC1+ sorted populations.</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Adult stem cells", "Stem cell lines", "Cell differentiation", "morphogenesis", "histology", "Molecular cell biology", "Cellular types", "Epithelial cells", "Cell growth", "derived", "progenitors", "mammoplasties", "hollow", "structures", "luminal-", "myoepithelial-like"], "article_id"=>711573, "categories"=>["Biological Sciences"], "users"=>["Chanel E. Smart", "Brian J. Morrison", "Jodi M. Saunus", "Ana Cristina Vargas", "Patricia Keith", "Lynne Reid", "Leesa Wockner", "Marjan Askarian Amiri", "Debina Sarkar", "Peter T. Simpson", "Catherine Clarke", "Chris W. Schmidt", "Brent A. Reynolds", "Sunil R. Lakhani", "J. Alejandro Lopez"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0064388.g005", "stats"=>{"downloads"=>3, "page_views"=>83, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mammospheres_derived_from_MUC1_or_CD10_progenitors_from_reduction_mammoplasties_comprise_hollow_and_solid_structures_consistent_with_luminal_and_myoepithelial_like_morphologies_/711573", "title"=>"Mammospheres derived from MUC1+ or CD10+ progenitors from reduction mammoplasties comprise hollow and solid structures consistent with luminal- and myoepithelial-like morphologies.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-04 00:26:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/1074325"], "description"=>"<p>MUC1<sup>+</sup> sphere cultures were prepared as described in Fig. 5A, in parallel with matched MUC1<sup>+</sup> adherent cultures, then dissociated and used in a colony forming cell (CFC) assay to determine the clonogenicity of luminal and myoepithelial/bipotent progenitor compartments (which give rise to colonies with luminal and mixed/myo morphologies respectively). (A) <b>Light micrographs of Geimsa-stained colonies</b> grown from dissociated spheres and parallel adherent cultures. Representative colonies are shown for adherent- (i-iii) and sphere-derived (iv-vi) cells. This includes examples of luminal (i, iv and v), mixed (ii, iii) and myoepithelial (vi) colony morphologies. (B) <b>Overall clonogenicity</b> (colonies formed as a percentage of total cells seeded) of spheres and parallel adherent cultures. (C) <b>Comparison of luminal and myo/bipotent progenitor cell clonogenicity of spheres and parallel adherent cultures.</b> Data are from four biological replicates (specimens from 4 patients), each performed in triplicate. Statistical tests used were paired, two-tailed students t-tests (<i>P</i> values indicated; <i>ns</i> = not significant).</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Adult stem cells", "Stem cell lines", "Cell differentiation", "morphogenesis", "histology", "Molecular cell biology", "Cellular types", "Epithelial cells", "Cell growth", "frequencies", "luminal", "mammary", "epithelial", "cells", "mammosphere"], "article_id"=>711576, "categories"=>["Biological Sciences"], "users"=>["Chanel E. Smart", "Brian J. Morrison", "Jodi M. Saunus", "Ana Cristina Vargas", "Patricia Keith", "Lynne Reid", "Leesa Wockner", "Marjan Askarian Amiri", "Debina Sarkar", "Peter T. Simpson", "Catherine Clarke", "Chris W. Schmidt", "Brent A. Reynolds", "Sunil R. Lakhani", "J. Alejandro Lopez"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0064388.g006", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Relative_frequencies_of_luminal_and_myoepithelial_bipotent_human_mammary_epithelial_cells_in_MUC1_derived_mammosphere_cultures_/711576", "title"=>"Relative frequencies of luminal and myoepithelial/bipotent human mammary epithelial cells in MUC1<sup>+</sup>-derived mammosphere cultures.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-04 00:26:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/1074327"], "description"=>"<p>IHC methods.</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Adult stem cells", "Stem cell lines", "Cell differentiation", "morphogenesis", "histology", "Molecular cell biology", "Cellular types", "Epithelial cells", "Cell growth"], "article_id"=>711578, "categories"=>["Biological Sciences"], "users"=>["Chanel E. Smart", "Brian J. Morrison", "Jodi M. Saunus", "Ana Cristina Vargas", "Patricia Keith", "Lynne Reid", "Leesa Wockner", "Marjan Askarian Amiri", "Debina Sarkar", "Peter T. Simpson", "Catherine Clarke", "Chris W. Schmidt", "Brent A. Reynolds", "Sunil R. Lakhani", "J. Alejandro Lopez"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0064388.t001", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_IHC_methods_/711578", "title"=>"IHC methods.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-06-04 00:26:18"}
  • {"files"=>["https://ndownloader.figshare.com/files/1074330", "https://ndownloader.figshare.com/files/1074334", "https://ndownloader.figshare.com/files/1074347", "https://ndownloader.figshare.com/files/1074349", "https://ndownloader.figshare.com/files/1074357", "https://ndownloader.figshare.com/files/1074359"], "description"=>"<div><p>Mammosphere and breast tumoursphere culture have gained popularity as <i>in vitro</i> assays for propagating and analysing normal and cancer stem cells. Whether the spheres derived from different sources or parent cultures themselves are indeed single entities enriched in stem/progenitor cells compared to other culture formats has not been fully determined. We surveyed sphere-forming capacity across 26 breast cell lines, immunophenotyped spheres from six luminal- and basal-like lines by immunohistochemistry and flow cytometry and compared clonogenicity between sphere, adherent and matrigel culture formats using <i>in vitro</i> functional assays. Analyses revealed morphological and molecular intra- and inter-sphere heterogeneity, consistent with adherent parental cell line phenotypes. Flow cytometry showed sphere culture does not universally enrich for markers previously associated with stem cell phenotypes, although we found some cell-line specific changes between sphere and adherent formats. Sphere-forming efficiency was significantly lower than adherent or matrigel clonogenicity and constant over serial passage. Surprisingly, self-renewal capacity of sphere-derived cells was similar/lower than other culture formats. We observed significant correlation between long-term-proliferating-cell symmetric division rates in sphere and adherent cultures, suggesting functional overlap between the compartments sustaining them. Experiments with normal primary human mammary epithelia, including sorted luminal (MUC1<sup>+</sup>) and basal/myoepithelial (CD10<sup>+</sup>) cells revealed distinct luminal-like, basal-like and mesenchymal entities amongst primary mammospheres. Morphological and colony-forming-cell assay data suggested mammosphere culture may enrich for a luminal progenitor phenotype, or induce reversion/relaxation of the basal/mesenchymal <i>in vitro</i> selection occurring with adherent culture. Overall, cell line tumourspheres and primary mammospheres are not homogenous entities enriched for stem cells, suggesting a more cautious approach to interpreting data from these assays and careful consideration of its limitations. Sphere culture may represent an alternative 3-dimensional culture system which rather than universally ‘enriching’ for stem cells, has utility as one of a suite of functional assays that provide a read-out of progenitor activity.</p></div>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Adult stem cells", "Stem cell lines", "Cell differentiation", "morphogenesis", "histology", "Molecular cell biology", "Cellular types", "Epithelial cells", "Cell growth", "cancer", "tumourspheres", "epithelia", "mammospheres", "demonstrates", "inter-", "intrasphere", "heterogeneity"], "article_id"=>711581, "categories"=>["Biological Sciences"], "users"=>["Chanel E. Smart", "Brian J. Morrison", "Jodi M. Saunus", "Ana Cristina Vargas", "Patricia Keith", "Lynne Reid", "Leesa Wockner", "Marjan Askarian Amiri", "Debina Sarkar", "Peter T. Simpson", "Catherine Clarke", "Chris W. Schmidt", "Brent A. Reynolds", "Sunil R. Lakhani", "J. Alejandro Lopez"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0064388.s001", "https://dx.doi.org/10.1371/journal.pone.0064388.s002", "https://dx.doi.org/10.1371/journal.pone.0064388.s003", "https://dx.doi.org/10.1371/journal.pone.0064388.s004", "https://dx.doi.org/10.1371/journal.pone.0064388.s005", "https://dx.doi.org/10.1371/journal.pone.0064388.s006"], "stats"=>{"downloads"=>39, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_Vitro_Analysis_of_Breast_Cancer_Cell_Line_Tumourspheres_and_Primary_Human_Breast_Epithelia_Mammospheres_Demonstrates_Inter_and_Intrasphere_Heterogeneity/711581", "title"=>"<i>In Vitro</i> Analysis of Breast Cancer Cell Line Tumourspheres and Primary Human Breast Epithelia Mammospheres Demonstrates Inter- and Intrasphere Heterogeneity", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-06-04 00:26:21"}

PMC Usage Stats | Further Information

  • {"unique-ip"=>"45", "full-text"=>"34", "pdf"=>"20", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"37", "supp-data"=>"4", "cited-by"=>"0", "year"=>"2013", "month"=>"6"}
  • {"unique-ip"=>"61", "full-text"=>"65", "pdf"=>"47", "abstract"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"25", "supp-data"=>"3", "cited-by"=>"0", "year"=>"2013", "month"=>"7"}
  • {"unique-ip"=>"48", "full-text"=>"49", "pdf"=>"33", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"32", "supp-data"=>"6", "cited-by"=>"0", "year"=>"2013", "month"=>"8"}
  • {"unique-ip"=>"60", "full-text"=>"75", "pdf"=>"34", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"7", "supp-data"=>"7", "cited-by"=>"0", "year"=>"2013", "month"=>"9"}
  • {"unique-ip"=>"28", "full-text"=>"33", "pdf"=>"21", "abstract"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"7", "cited-by"=>"0", "year"=>"2013", "month"=>"10"}
  • {"unique-ip"=>"28", "full-text"=>"24", "pdf"=>"10", "abstract"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"10", "supp-data"=>"3", "cited-by"=>"0", "year"=>"2013", "month"=>"11"}
  • {"unique-ip"=>"23", "full-text"=>"25", "pdf"=>"11", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"15", "supp-data"=>"5", "cited-by"=>"0", "year"=>"2013", "month"=>"12"}
  • {"unique-ip"=>"34", "full-text"=>"29", "pdf"=>"10", "abstract"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"23", "supp-data"=>"3", "cited-by"=>"0", "year"=>"2014", "month"=>"1"}
  • {"unique-ip"=>"18", "full-text"=>"15", "pdf"=>"7", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"6", "supp-data"=>"2", "cited-by"=>"0", "year"=>"2014", "month"=>"2"}
  • {"unique-ip"=>"28", "full-text"=>"27", "pdf"=>"17", "abstract"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"9", "supp-data"=>"3", "cited-by"=>"0", "year"=>"2014", "month"=>"3"}
  • {"unique-ip"=>"19", "full-text"=>"16", "pdf"=>"7", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"19", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"5"}
  • {"unique-ip"=>"21", "full-text"=>"20", "pdf"=>"9", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"7", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"6"}
  • {"unique-ip"=>"19", "full-text"=>"20", "pdf"=>"5", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"6", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"4"}
  • {"unique-ip"=>"28", "full-text"=>"34", "pdf"=>"11", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"3", "cited-by"=>"0", "year"=>"2015", "month"=>"4"}
  • {"unique-ip"=>"27", "full-text"=>"24", "pdf"=>"16", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"5"}
  • {"unique-ip"=>"21", "full-text"=>"20", "pdf"=>"11", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"6"}
  • {"unique-ip"=>"22", "full-text"=>"19", "pdf"=>"10", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"13", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"7"}
  • {"unique-ip"=>"32", "full-text"=>"47", "pdf"=>"18", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"9", "supp-data"=>"4", "cited-by"=>"0", "year"=>"2015", "month"=>"3"}
  • {"unique-ip"=>"25", "full-text"=>"27", "pdf"=>"11", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"13", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"2"}
  • {"unique-ip"=>"12", "full-text"=>"12", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"8"}
  • {"unique-ip"=>"22", "full-text"=>"21", "pdf"=>"8", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"6", "supp-data"=>"3", "cited-by"=>"0", "year"=>"2015", "month"=>"9"}
  • {"unique-ip"=>"20", "full-text"=>"20", "pdf"=>"6", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"2", "cited-by"=>"0", "year"=>"2015", "month"=>"10"}
  • {"unique-ip"=>"19", "full-text"=>"20", "pdf"=>"9", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"11", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"7"}
  • {"unique-ip"=>"10", "full-text"=>"9", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"8"}
  • {"unique-ip"=>"23", "full-text"=>"12", "pdf"=>"9", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"7", "cited-by"=>"0", "year"=>"2014", "month"=>"9"}
  • {"unique-ip"=>"29", "full-text"=>"21", "pdf"=>"12", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"11", "supp-data"=>"3", "cited-by"=>"1", "year"=>"2014", "month"=>"10"}
  • {"unique-ip"=>"21", "full-text"=>"25", "pdf"=>"9", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"2"}
  • {"unique-ip"=>"12", "full-text"=>"11", "pdf"=>"9", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2014", "month"=>"11"}
  • {"unique-ip"=>"14", "full-text"=>"17", "pdf"=>"11", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"5", "cited-by"=>"0", "year"=>"2014", "month"=>"12"}
  • {"unique-ip"=>"26", "full-text"=>"25", "pdf"=>"17", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"1"}
  • {"unique-ip"=>"27", "full-text"=>"20", "pdf"=>"14", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"10", "supp-data"=>"1", "cited-by"=>"2", "year"=>"2015", "month"=>"11"}
  • {"unique-ip"=>"19", "full-text"=>"18", "pdf"=>"8", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"4", "cited-by"=>"0", "year"=>"2015", "month"=>"12"}
  • {"unique-ip"=>"13", "full-text"=>"12", "pdf"=>"6", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"1"}
  • {"unique-ip"=>"22", "full-text"=>"19", "pdf"=>"9", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2016", "month"=>"3"}
  • {"unique-ip"=>"25", "full-text"=>"21", "pdf"=>"10", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"9", "supp-data"=>"3", "cited-by"=>"0", "year"=>"2016", "month"=>"4"}
  • {"unique-ip"=>"24", "full-text"=>"20", "pdf"=>"11", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"6", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"5"}
  • {"unique-ip"=>"12", "full-text"=>"16", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"3", "cited-by"=>"0", "year"=>"2016", "month"=>"6"}
  • {"unique-ip"=>"21", "full-text"=>"19", "pdf"=>"6", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2016", "month"=>"7"}
  • {"unique-ip"=>"20", "full-text"=>"19", "pdf"=>"8", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"8"}
  • {"unique-ip"=>"21", "full-text"=>"24", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"9"}
  • {"unique-ip"=>"31", "full-text"=>"29", "pdf"=>"9", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"10"}
  • {"unique-ip"=>"23", "full-text"=>"20", "pdf"=>"5", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"8", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2016", "month"=>"11"}
  • {"unique-ip"=>"20", "full-text"=>"31", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"12"}
  • {"unique-ip"=>"25", "full-text"=>"31", "pdf"=>"5", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"1"}
  • {"unique-ip"=>"24", "full-text"=>"17", "pdf"=>"6", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"15", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"2"}
  • {"unique-ip"=>"23", "full-text"=>"23", "pdf"=>"9", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"6", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2017", "month"=>"3"}
  • {"unique-ip"=>"22", "full-text"=>"25", "pdf"=>"8", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"4"}
  • {"unique-ip"=>"34", "full-text"=>"41", "pdf"=>"10", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"3", "cited-by"=>"0", "year"=>"2017", "month"=>"5"}
  • {"unique-ip"=>"28", "full-text"=>"27", "pdf"=>"7", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"8", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"6"}
  • {"unique-ip"=>"25", "full-text"=>"29", "pdf"=>"9", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"6", "supp-data"=>"3", "cited-by"=>"0", "year"=>"2017", "month"=>"7"}
  • {"unique-ip"=>"28", "full-text"=>"29", "pdf"=>"10", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"8"}
  • {"unique-ip"=>"17", "full-text"=>"19", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"9"}
  • {"unique-ip"=>"42", "full-text"=>"43", "pdf"=>"13", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"7", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2017", "month"=>"10"}
  • {"unique-ip"=>"36", "full-text"=>"37", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"7", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"11"}
  • {"unique-ip"=>"20", "full-text"=>"14", "pdf"=>"5", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"12", "cited-by"=>"0", "year"=>"2017", "month"=>"12"}
  • {"unique-ip"=>"29", "full-text"=>"30", "pdf"=>"11", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"3", "cited-by"=>"0", "year"=>"2018", "month"=>"1"}
  • {"unique-ip"=>"1", "full-text"=>"1", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"2"}
  • {"unique-ip"=>"37", "full-text"=>"38", "pdf"=>"7", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"12", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2018", "month"=>"3"}
  • {"unique-ip"=>"30", "full-text"=>"32", "pdf"=>"5", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"5", "cited-by"=>"0", "year"=>"2019", "month"=>"1"}
  • {"unique-ip"=>"37", "full-text"=>"41", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"6", "cited-by"=>"0", "year"=>"2018", "month"=>"5"}
  • {"unique-ip"=>"25", "full-text"=>"31", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"6"}
  • {"unique-ip"=>"30", "full-text"=>"31", "pdf"=>"10", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2018", "month"=>"8"}
  • {"unique-ip"=>"33", "full-text"=>"40", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2018", "month"=>"10"}
  • {"unique-ip"=>"29", "full-text"=>"28", "pdf"=>"6", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"3", "cited-by"=>"0", "year"=>"2018", "month"=>"4"}
  • {"unique-ip"=>"30", "full-text"=>"27", "pdf"=>"8", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"5", "cited-by"=>"0", "year"=>"2018", "month"=>"7"}
  • {"unique-ip"=>"20", "full-text"=>"22", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"12"}
  • {"unique-ip"=>"38", "full-text"=>"37", "pdf"=>"6", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"8", "cited-by"=>"0", "year"=>"2018", "month"=>"11"}
  • {"unique-ip"=>"37", "full-text"=>"36", "pdf"=>"12", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"7", "cited-by"=>"0", "year"=>"2018", "month"=>"9"}
  • {"unique-ip"=>"29", "full-text"=>"27", "pdf"=>"7", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"2", "cited-by"=>"0", "year"=>"2019", "month"=>"2"}
  • {"unique-ip"=>"33", "full-text"=>"36", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"7", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"3"}
  • {"unique-ip"=>"39", "full-text"=>"43", "pdf"=>"6", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"4"}
  • {"unique-ip"=>"52", "full-text"=>"56", "pdf"=>"13", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"5", "cited-by"=>"0", "year"=>"2019", "month"=>"5"}
  • {"unique-ip"=>"39", "full-text"=>"35", "pdf"=>"17", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"7", "cited-by"=>"0", "year"=>"2019", "month"=>"8"}
  • {"unique-ip"=>"42", "full-text"=>"47", "pdf"=>"7", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"4", "cited-by"=>"0", "year"=>"2019", "month"=>"9"}
  • {"unique-ip"=>"34", "full-text"=>"30", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"10"}
  • {"unique-ip"=>"27", "full-text"=>"26", "pdf"=>"9", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"12", "cited-by"=>"0", "year"=>"2019", "month"=>"12"}
  • {"unique-ip"=>"34", "full-text"=>"37", "pdf"=>"7", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"26", "cited-by"=>"0", "year"=>"2020", "month"=>"2"}
  • {"unique-ip"=>"24", "full-text"=>"25", "pdf"=>"6", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"2", "cited-by"=>"0", "year"=>"2020", "month"=>"3"}
  • {"unique-ip"=>"12", "full-text"=>"11", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"4"}
  • {"unique-ip"=>"13", "full-text"=>"14", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2020", "month"=>"5"}
  • {"unique-ip"=>"29", "full-text"=>"29", "pdf"=>"9", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"6"}
  • {"unique-ip"=>"19", "full-text"=>"21", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"7"}
  • {"unique-ip"=>"20", "full-text"=>"18", "pdf"=>"5", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"8", "cited-by"=>"0", "year"=>"2020", "month"=>"8"}
  • {"unique-ip"=>"20", "full-text"=>"24", "pdf"=>"5", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2020", "month"=>"9"}
  • {"unique-ip"=>"33", "full-text"=>"36", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"10"}

Relative Metric

{"start_date"=>"2013-01-01T00:00:00Z", "end_date"=>"2013-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences", "average_usage"=>[269, 466, 588, 697, 800, 896, 988, 1076, 1165, 1254, 1340, 1417]}, {"subject_area"=>"/Biology and life sciences/Developmental biology", "average_usage"=>[275, 472, 605, 720, 822, 921, 1013, 1106, 1200, 1289, 1378, 1459, 1531]}, {"subject_area"=>"/Medicine and health sciences/Oncology", "average_usage"=>[249, 468, 599, 718, 820, 920, 1008, 1093, 1181, 1281, 1357, 1444, 1517]}]}
Loading … Spinner
There are currently no alerts