In Vitro Megakaryocyte Differentiation and Proplatelet Formation in Ph-Negative Classical Myeloproliferative Neoplasms: Distinct Patterns in the Different Clinical Phenotypes
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{"title"=>"In vitro megakaryocyte differentiation and proplatelet formation in Ph-negative classical myeloproliferative neoplasms: Distinct patterns in the different clinical phenotypes", "type"=>"journal", "authors"=>[{"first_name"=>"Alessandra", "last_name"=>"Balduini", "scopus_author_id"=>"6508339245"}, {"first_name"=>"Stefania", "last_name"=>"Badalucco", "scopus_author_id"=>"26027756600"}, {"first_name"=>"Maria Teresa", "last_name"=>"Pugliano", "scopus_author_id"=>"26642692400"}, {"first_name"=>"Denis", "last_name"=>"Baev", "scopus_author_id"=>"24512138100"}, {"first_name"=>"Annalisa", "last_name"=>"de Silvestri", "scopus_author_id"=>"6701866631"}, {"first_name"=>"Marco", "last_name"=>"Cattaneo", "scopus_author_id"=>"57189455495"}, {"first_name"=>"Vittorio", "last_name"=>"Rosti", "scopus_author_id"=>"7004396919"}, {"first_name"=>"Giovanni", "last_name"=>"Barosi", "scopus_author_id"=>"7006254277"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"79959254853", "pmid"=>"21698292", "scopus"=>"2-s2.0-79959254853", "pui"=>"361955749", "isbn"=>"1932-6203 (Electronic)\\n1932-6203 (Linking)", "issn"=>"19326203", "doi"=>"10.1371/journal.pone.0021015"}, "id"=>"e3b6f71f-b232-3609-ab7f-ebfe808902d1", "abstract"=>"BACKGROUND Ph-negative myeloproliferative neoplasms (MPNs) are clonal disorders that include primary myelofibrosis (PMF), polycythemia vera (PV) and essential thrombocythemia (ET). Although the pathogenesis of MPNs is still incompletely understood, an involvement of the megakaryocyte lineage is a distinctive feature. METHODOLOGY/PRINCIPAL FINDINGS We analyzed the in vitro megakaryocyte differentiation and proplatelet formation in 30 PMF, 8 ET, 8 PV patients, and 17 healthy controls (CTRL). Megakaryocytes were differentiated from peripheral blood CD34(+) or CD45(+) cells in the presence of thrombopoietin. Megakaryocyte output was higher in MPN patients than in CTRL with no correlation with the JAK2 V617F mutation. PMF-derived megakaryocytes displayed nuclei with a bulbous appearance, were smaller than ET- or PV-derived megakaryocytes and formed proplatelets that presented several structural alterations. In contrast, ET- and PV-derived megakaryocytes produced more proplatelets with a striking increase in bifurcations and tips compared to both control and PMF. Proplatelets formation was correlated with platelet counts in patient peripheral blood. Patients with pre-fibrotic PMF had a pattern of megakaryocyte proliferation and proplatelet formation that was similar to that of fibrotic PMF and different from that of ET. CONCLUSIONS/SIGNIFICANCE In conclusion, MPNs are associated with high megakaryocyte proliferative potential. Profound differences in megakaryocyte morphology and proplatelet formation distinguish PMF, both fibrotic and prefibrotic, from ET and PV.", "link"=>"http://www.mendeley.com/research/vitro-megakaryocyte-differentiation-proplatelet-formation-phnegative-classical-myeloproliferative-ne", "reader_count"=>43, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>1, "Researcher"=>10, "Student > Doctoral Student"=>5, "Student > Ph. D. Student"=>17, "Student > Postgraduate"=>1, "Student > Master"=>6, "Student > Bachelor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>1, "Researcher"=>10, "Student > Doctoral Student"=>5, "Student > Ph. D. Student"=>17, "Student > Postgraduate"=>1, "Student > Master"=>6, "Student > Bachelor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>8, "Agricultural and Biological Sciences"=>22, "Medicine and Dentistry"=>11}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>11}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>22}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>8}, "Unspecified"=>{"Unspecified"=>2}}, "reader_count_by_country"=>{"Belgium"=>1, "Japan"=>2, "France"=>1}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/765673"], "description"=>"<p>CD45<sup>+</sup> (A) and CD34<sup>+</sup> (B) cells from peripheral blood were sorted as described in Methods and cultured for 14 days. At the end of the cultures, the yield of megakaryocyte was calculated as output of CD41<sup>+</sup> cells with megakaryocyte morphology. CTRL: healthy controls; PMF: primary myeolofibrosis; ET: essential thrombocytemia; PV polycytemia vera. *p<0.01.</p>", "links"=>[], "tags"=>["plots", "megakaryocyte", "suspension"], "article_id"=>436033, "categories"=>["Physiology", "Cancer", "Cell Biology", "Hematology", "Immunology"], "users"=>["Alessandra Balduini", "Stefania Badalucco", "Maria Teresa Pugliano", "Denis Baev", "Annalisa De Silvestri", "Marco Cattaneo", "Vittorio Rosti", "Giovanni Barosi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021015.g001", "stats"=>{"downloads"=>2, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Box_and_whisker_plots_of_megakaryocyte_output_in_suspension_cultures_/436033", "title"=>"Box-and-whisker plots of megakaryocyte output in suspension cultures.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 17:33:51"}
  • {"files"=>["https://ndownloader.figshare.com/files/766275"], "description"=>"<p>(A) Box-and-whisker plots of proplatelet output, expressed as percent of total megakaryocytes, *p<0.01. (B) Representative picture of differently organized proplatelet revealed by alpha tubulin staining (green) of controls (I), PMF (II), ET (III) and PV (IV). Nuclei are counterstained with Hoechst 33258 (blu). Scale bars are 15 µm.</p>", "links"=>[], "tags"=>["megakaryocytes", "adhering"], "article_id"=>436643, "categories"=>["Physiology", "Cancer", "Cell Biology", "Hematology", "Immunology"], "users"=>["Alessandra Balduini", "Stefania Badalucco", "Maria Teresa Pugliano", "Denis Baev", "Annalisa De Silvestri", "Marco Cattaneo", "Vittorio Rosti", "Giovanni Barosi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021015.g005", "stats"=>{"downloads"=>1, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Proplatelet_formation_by_megakaryocytes_adhering_to_fibrinogen_/436643", "title"=>"Proplatelet formation by megakaryocytes adhering to fibrinogen.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 17:37:11"}
  • {"files"=>["https://ndownloader.figshare.com/files/766417"], "description"=>"<p>*Age, Hb, Hct, WBC, and Plt counts are expressed as median (range). Diagnosis was made according to WHO criteria (see text for details). Values reported in the table are those at the moment in which blood was drawn for experiments. PMF and ET patients were at diagnosis or before the beginning of any cytoreductive therapy. Patients with PV were all under treatment with phlebotomy (but not with cytostatic therapy) in order to try to lower their Hct below 45% (males) or 42% (females). Healthy CTRLs hematological values were assessed before treatment with G-CSF (see text for details).</p>", "links"=>[], "tags"=>["immunology", "physiology", "cell biology", "hematology", "oncology"], "article_id"=>436774, "categories"=>["Physiology", "Cancer", "Cell Biology", "Hematology", "Immunology"], "users"=>["Alessandra Balduini", "Stefania Badalucco", "Maria Teresa Pugliano", "Denis Baev", "Annalisa De Silvestri", "Marco Cattaneo", "Vittorio Rosti", "Giovanni Barosi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021015.t001", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Epidemiological_and_clinical_data_of_patient_and_control_populations_/436774", "title"=>"Epidemiological and clinical data of patient and control populations<sup>*</sup>.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 17:37:52"}
  • {"files"=>["https://ndownloader.figshare.com/files/765863"], "description"=>"<p>(A) Representative picture of differently shaped CD41<sup>+</sup> (red) cells of controls (I), pre-fibrotic PMF(II), PMF (III), ET (IV) and PV (V). Nuclei are counterstained with Hoechst 33258 (blue). Scale bars are 15 µm. (B) Ploidy was analyzed as described in Methods. PMF derived megakaryocytes presented lower polyploidy with respect to controls. (C) PMF derived CD41<sup>+</sup> megakaryocytes (red) displayed bulbous and hypo-segmented nuclei.(blue) (left panel). Scale bar is 15 µm. Means ± SD of the percentage of megakaryocytes displaying abnormal nuclei in pre-fibrotic PMF and PMF, as compared to control, are reported in the right panel. (D) Diameters of MPN derived CD41<sup>+</sup> megakaryocytes (red) were analyzed as shown in the left panel and were performed as described in Methods. Scale bar is 10 µm. For each MPN category at least 100 megakaryocytes were analyzed, the means ± SD of diameters is reported in the right panel. *p<0.05.</p>", "links"=>[], "tags"=>["megakaryocyte", "morphology", "mpns", "controls"], "article_id"=>436225, "categories"=>["Physiology", "Cancer", "Cell Biology", "Hematology", "Immunology"], "users"=>["Alessandra Balduini", "Stefania Badalucco", "Maria Teresa Pugliano", "Denis Baev", "Annalisa De Silvestri", "Marco Cattaneo", "Vittorio Rosti", "Giovanni Barosi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021015.g002", "stats"=>{"downloads"=>0, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Characteristics_of_megakaryocyte_morphology_in_MPNs_and_controls_CTRL_/436225", "title"=>"Characteristics of megakaryocyte morphology in MPNs and controls (CTRL).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 17:34:56"}
  • {"files"=>["https://ndownloader.figshare.com/files/766125"], "description"=>"<p>(A) Representative picture of differently organized proplatelet revealed by alpha tubulin staining (green) of controls (I), PMF (II–III), ET (IV) and PV (V). Nuclei are counterstained with Hoechst 33258 (blu). Scale bars are 15 µm. (B) Proplatelet bifurcations (means ± SD), which were identified upon immunostaining with an antibody against alpha tubulin. (C) Proplatelet tips (means ± SD), which were identified as coiled coil formations localized at the end of proplatelet branches. *p<0.05.</p>", "links"=>[], "tags"=>["proplatelet"], "article_id"=>436490, "categories"=>["Physiology", "Cancer", "Cell Biology", "Hematology", "Immunology"], "users"=>["Alessandra Balduini", "Stefania Badalucco", "Maria Teresa Pugliano", "Denis Baev", "Annalisa De Silvestri", "Marco Cattaneo", "Vittorio Rosti", "Giovanni Barosi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021015.g004", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Analysis_of_proplatelet_architecture_/436490", "title"=>"Analysis of proplatelet architecture.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 17:36:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/765985"], "description"=>"<p>(A) Box-and-whisker plots of proplatelet output, expressed as percent of total megakaryocytes. (B) Correlation between the platelet count in peripheral blood and the number of proplatelets formed in culture in MPN patients. *p<0.01.</p>", "links"=>[], "tags"=>["differences", "megakaryocyte"], "article_id"=>436351, "categories"=>["Physiology", "Cancer", "Cell Biology", "Hematology", "Immunology"], "users"=>["Alessandra Balduini", "Stefania Badalucco", "Maria Teresa Pugliano", "Denis Baev", "Annalisa De Silvestri", "Marco Cattaneo", "Vittorio Rosti", "Giovanni Barosi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021015.g003", "stats"=>{"downloads"=>1, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MPNs_show_important_differences_in_megakaryocyte_function_and_PPF_/436351", "title"=>"MPNs show important differences in megakaryocyte function and PPF.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 17:35:38"}

PMC Usage Stats | Further Information

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Relative Metric

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