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{"title"=>"Adult human biliary tree stem cells differentiate to β-pancreatic islet cells by treatment with a recombinant human Pdx1 peptide", "type"=>"journal", "authors"=>[{"first_name"=>"Vincenzo", "last_name"=>"Cardinale", "scopus_author_id"=>"23569414600"}, {"first_name"=>"Rosa", "last_name"=>"Puca", "scopus_author_id"=>"17344525100"}, {"first_name"=>"Guido", "last_name"=>"Carpino", "scopus_author_id"=>"6508122425"}, {"first_name"=>"Gaia", "last_name"=>"Scafetta", "scopus_author_id"=>"55335977700"}, {"first_name"=>"Anastasia", "last_name"=>"Renzi", "scopus_author_id"=>"7006986165"}, {"first_name"=>"Michele", "last_name"=>"De Canio", "scopus_author_id"=>"6506901929"}, {"first_name"=>"Francesca", "last_name"=>"Sicilia", "scopus_author_id"=>"57194630147"}, {"first_name"=>"Lorenzo", "last_name"=>"Nevi", "scopus_author_id"=>"56866411600"}, {"first_name"=>"Domenico", "last_name"=>"Casa", "scopus_author_id"=>"56865693500"}, {"first_name"=>"Rocco", "last_name"=>"Panetta", "scopus_author_id"=>"56865960700"}, {"first_name"=>"Pasquale Bartolomeo", "last_name"=>"Berloco", "scopus_author_id"=>"7005248713"}, {"first_name"=>"Lola M.", "last_name"=>"Reid", "scopus_author_id"=>"7201752824"}, {"first_name"=>"Giorgio", "last_name"=>"Federici", "scopus_author_id"=>"7101698480"}, {"first_name"=>"Eugenio", "last_name"=>"Gaudio", "scopus_author_id"=>"7006542760"}, {"first_name"=>"Marella", "last_name"=>"Maroder", "scopus_author_id"=>"6603896568"}, {"first_name"=>"Domenico", "last_name"=>"Alvaro", "scopus_author_id"=>"7006353760"}], "year"=>2015, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84942155078", "pui"=>"606057532", "scopus"=>"2-s2.0-84942155078", "doi"=>"10.1371/journal.pone.0134677", "issn"=>"19326203"}, "id"=>"ea5fd13b-da12-3318-b929-56c5a944da38", "link"=>"http://www.mendeley.com/research/adult-human-biliary-tree-stem-cells-differentiate-%CE%B2pancreatic-islet-cells-treatment-recombinant-huma-1", "reader_count"=>11, "reader_count_by_academic_status"=>{"Researcher"=>3, "Student > Ph. D. Student"=>2, "Student > Postgraduate"=>1, "Student > Master"=>1, "Student > Bachelor"=>1, "Professor"=>1, "Unspecified"=>2}, "reader_count_by_user_role"=>{"Researcher"=>3, "Student > Ph. D. Student"=>2, "Student > Postgraduate"=>1, "Student > Master"=>1, "Student > Bachelor"=>1, "Professor"=>1, "Unspecified"=>2}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>4, "Medicine and Dentistry"=>1, "Chemistry"=>1, "Unspecified"=>2}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Chemistry"=>{"Chemistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>4}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>2}}, "reader_count_by_country"=>{"Mexico"=>1}, "group_count"=>2}

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• http://doi.org/10.1038/s41598-017-05858-0
• http://doi.org/10.1016/j.diff.2019.03.001
• http://doi.org/10.3389/fgene.2017.00075
• http://doi.org/10.3390/cells8080914
• http://doi.org/10.1007/s12015-020-10074-x
• http://doi.org/10.3390/cells8111443

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Figshare

• {"files"=>["https://ndownloader.figshare.com/files/2205662"], "description"=>"<p>(A) Clarified bacterial lysate was applied to a 10 mL column of Chelating Sepharose activated with Ni²⁺ ions. Chromatographic steps were followed by monitoring the absorbance at 280 nm. (B) The inset shows SDS-PAGE analysis of lysate sample [<i>S</i>], flow-through [<i>F</i>], wash [<i>W</i>] and elution [<i>E</i>] peak fractions. Molecular weight marker [<i>M</i>] was run in parallel. The purity was estimated by image analysis of SDS-polyacrylamide gels: the 43kDa recombinant protein is > 90% of the total protein content. The final yield was estimated at around 10 mg per liter of bacterial culture. (C) Identity of purified protein was checked by MALDI-TOF-TOF Mass Spectrometry. Peptide Mass Fingerprinting analysis showed a profile with 7 peptide tryptic masses matching with the theoretical masses originated by <i>in silico</i> digestion of human PDX1 sequence.</p>", "links"=>[], "tags"=>["hepatocyte cell line", "Adult Human Biliary Tree Stem Cells Differentiate", "Pdx 1 exposure", "hBTSC", "Pdx 1 protein", "pdx", "Pdx 1", "HepG 2 cell line", "time polymerase chain reaction", "pancreatic islet cells", "Pdx 1 peptide", "Recombinant Human Pdx 1 Peptide Generation"], "article_id"=>1505885, "categories"=>["Biological Sciences"], "users"=>["Vincenzo Cardinale", "Rosa Puca", "Guido Carpino", "Gaia Scafetta", "Anastasia Renzi", "Michele De Canio", "Francesca Sicilia", "Lorenzo Nevi", "Domenico Casa", "Rocco Panetta", "Pasquale Bartolomeo Berloco", "Lola M. Reid", "Giorgio Federici", "Eugenio Gaudio", "Marella Maroder", "Domenico Alvaro"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0134677.g001", "stats"=>{"downloads"=>1, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Nickel_Chelating_Sepharose_chromatography_of_recombinant_6His_tagged_Pdx1_and_identity_of_purified_Pdx1_protein_by_Mass_Spectrometry_/1505885", "title"=>"Nickel Chelating Sepharose chromatography of recombinant 6His-tagged Pdx1 and identity of purified Pdx1 protein by Mass Spectrometry.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-08-07 03:32:39"}
• {"files"=>["https://ndownloader.figshare.com/files/2205673"], "description"=>"<p>(A) In HepG2 cells the insulin mRNA level was not affected by 14 days of 0.1 μM Pdx1 administration, while it increased significantly (average 6 folds) in hBTSCs cultured for 14 days in Kubota’s Medium (KM) containing 0.1 μM Pdx1in comparison with control medium (KM) (data are means ± SD of 6 experiments; ***p< 0.001). The effect of Pdx1 administration on hBTSCs was similar to treatment with a Hormonally Defined Medium for Pancreatic islet cell differentiation (HDM-P). PANC-1 cells and pancreatic islets (islets) were used as positive controls. (B, C) In hBTSCs cultured for 14 days in KM containing 0.1 μM Pdx1, the glucagon and somatostatin mRNA levels increased 6- (data are means ± SD of 6 experiments; ***p< 0.001) and 2-folds (data are means ± SD of 6 experiments; **p< 0.01), respectively, in comparison with KM. (D) In hBTSCs cultured for 14 days in KM containing 0.1 μM Pdx1, MafA (4.5-folds) (data are means ± SD of 6 experiments; **p< 0.01) or Pdx1 (2-folds) (data are means ± SD of 6 experiments; **p< 0.01) gene expression increased in comparison with KM, while EpCAM mRNA decreased (average 4-folds) (data are means ± SD of 6 experiments; *p< 0.05).</p>", "links"=>[], "tags"=>["hepatocyte cell line", "Adult Human Biliary Tree Stem Cells Differentiate", "Pdx 1 exposure", "hBTSC", "Pdx 1 protein", "pdx", "Pdx 1", "HepG 2 cell line", "time polymerase chain reaction", "pancreatic islet cells", "Pdx 1 peptide", "Recombinant Human Pdx 1 Peptide Generation"], "article_id"=>1505891, "categories"=>["Biological Sciences"], "users"=>["Vincenzo Cardinale", "Rosa Puca", "Guido Carpino", "Gaia Scafetta", "Anastasia Renzi", "Michele De Canio", "Francesca Sicilia", "Lorenzo Nevi", "Domenico Casa", "Rocco Panetta", "Pasquale Bartolomeo Berloco", "Lola M. Reid", "Giorgio Federici", "Eugenio Gaudio", "Marella Maroder", "Domenico Alvaro"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0134677.g003", "stats"=>{"downloads"=>0, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Human_biliary_tree_stem_cell_hBTSC_differentiation_towards_pancreatic_fate_induced_in_vitro_by_Pdx1_peptide_/1505891", "title"=>"Human biliary tree stem cell (hBTSC) differentiation towards pancreatic fate induced <i>in vitro</i> by Pdx1 peptide.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-08-07 03:32:39"}
• {"files"=>["https://ndownloader.figshare.com/files/2205666"], "description"=>"<p>(A) hBTSCs and HepG2 were exposed to different concentrations of Pdx1 (0.1 μM or 0.5 μM). Incubation with 0.1 μM Pdx1 determined the preservation of cell viability. When cells were treated with 0.5 μM Pdx1, cell viability significantly decreased after 48 hours in both hBTSCs and HepG2 as observed by Trypan blue exclusion assay (data are means ± SD of 6 experiments; **p< 0.01; ***p< 0.001). (B) After 24 hours of treatment with 0.1 μM Pdx1 recombinant protein, Western Blot analyses showed the presence of Pdx1 recombinant protein (His-Tagged protein) in treated hBTSCs and HepG2. The densitometry histograms showed an equal amount of His-Tagged protein in hBTSCs and HepG2 cells treated with 0.1 μM Pdx1 (data are means ± SD of 3 experiments). (C) Immunofluorescence analysis showed the internalization and nuclear translocation of His-Tagged recombinant protein (red nuclei) in treated cells. Nuclei are displayed in blue (DAPI). Original Magnification = 20X left image or 40X right images.</p>", "links"=>[], "tags"=>["hepatocyte cell line", "Adult Human Biliary Tree Stem Cells Differentiate", "Pdx 1 exposure", "hBTSC", "Pdx 1 protein", "pdx", "Pdx 1", "HepG 2 cell line", "time polymerase chain reaction", "pancreatic islet cells", "Pdx 1 peptide", "Recombinant Human Pdx 1 Peptide Generation"], "article_id"=>1505889, "categories"=>["Biological Sciences"], "users"=>["Vincenzo Cardinale", "Rosa Puca", "Guido Carpino", "Gaia Scafetta", "Anastasia Renzi", "Michele De Canio", "Francesca Sicilia", "Lorenzo Nevi", "Domenico Casa", "Rocco Panetta", "Pasquale Bartolomeo Berloco", "Lola M. Reid", "Giorgio Federici", "Eugenio Gaudio", "Marella Maroder", "Domenico Alvaro"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0134677.g002", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Pdx1_effects_on_cell_viability_and_Pdx1_internalization_/1505889", "title"=>"Pdx1 effects on cell viability and Pdx1 internalization.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-08-07 03:32:39"}
• {"files"=>["https://ndownloader.figshare.com/files/2205680", "https://ndownloader.figshare.com/files/2205681"], "description"=>"<div><p>Generation of β-pancreatic cells represents a major goal in research. The aim of this study was to explore a protein-based strategy to induce differentiation of human biliary tree stem cells (hBTSCs) towards β-pancreatic cells. A plasmid containing the sequence of the human pancreatic and duodenal homeobox 1 (PDX1) has been expressed in E. coli. Epithelial-Cell-Adhesion-Molecule positive hBTSCs or mature human hepatocyte cell line, HepG2, were grown in medium to which Pdx1 peptide was added. Differentiation toward pancreatic islet cells were evaluated by the expression of the β-cell transcription factors, Pdx1 and musculoapo-neurotic fibrosarcoma oncogene homolog A, and of the pancreatic hormones, insulin, glucagon, and somatostatin, investigated by real time polymerase chain reaction, western blot, light microscopy and immunofluorescence. C-peptide secretion in response to high glucose was also measured. Results indicated how purified Pdx1 protein corresponding to the primary structure of the human Pdx1 by mass spectroscopy was efficiently produced in bacteria, and transduced into hBTSCs. Pdx1 exposure triggered the expression of both intermediate and mature stage β-cell differentiation markers only in hBTSCs but not in HepG2 cell line. Furthermore, hBTSCs exposed to Pdx1 showed up-regulation of insulin, glucagon and somatostatin genes and formation of 3-dimensional islet-like structures intensely positive for insulin and glucagon. Finally, Pdx1-induced islet-like structures exhibited glucose-regulated C-peptide secretion. In conclusion, the human Pdx1 is highly effective in triggering hBTSC differentiation toward functional β-pancreatic cells.</p></div>", "links"=>[], "tags"=>["hepatocyte cell line", "Adult Human Biliary Tree Stem Cells Differentiate", "Pdx 1 exposure", "hBTSC", "Pdx 1 protein", "pdx", "Pdx 1", "HepG 2 cell line", "time polymerase chain reaction", "pancreatic islet cells", "Pdx 1 peptide", "Recombinant Human Pdx 1 Peptide Generation"], "article_id"=>1505898, "categories"=>["Biological Sciences"], "users"=>["Vincenzo Cardinale", "Rosa Puca", "Guido Carpino", "Gaia Scafetta", "Anastasia Renzi", "Michele De Canio", "Francesca Sicilia", "Lorenzo Nevi", "Domenico Casa", "Rocco Panetta", "Pasquale Bartolomeo Berloco", "Lola M. Reid", "Giorgio Federici", "Eugenio Gaudio", "Marella Maroder", "Domenico Alvaro"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0134677.s001", "https://dx.doi.org/10.1371/journal.pone.0134677.s002"], "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Adult_Human_Biliary_Tree_Stem_Cells_Differentiate_to_946_Pancreatic_Islet_Cells_by_Treatment_with_a_Recombinant_Human_Pdx1_Peptide_/1505898", "title"=>"Adult Human Biliary Tree Stem Cells Differentiate to β-Pancreatic Islet Cells by Treatment with a Recombinant Human Pdx1 Peptide", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2015-08-07 03:32:39"}
• {"files"=>["https://ndownloader.figshare.com/files/2205675"], "description"=>"<p>(A) Morphologically (Phase Contrast: Ph-C; Original Magnification = 10X), after 14 days in basal condition, hBTSCs formed large colonies composed of small, densely packed, and uniform EpCAM-positive cells (Immunofluorescence for EpCAM; Original Magnification = 20X). After 14 days in cultures containing 0.1 μM Pdx1, several islet-like structures were present (Phase Contrast: Ph-C; Original Magnification = 10X); these structures were mostly composed of insulin-positive cells (Immunofluorescence for Insulin; Original Magnification = 10X); and some glucagon-positive cells were located at the periphery of these islet-like structures (double immunofluorescence for Insulin and Glucagon; Original Magnification = 20X). (B) Functionally, hBTSCs in Kubota’s Medium (KM) or in KM plus 0.1 μM Pdx1 were exposed to low (5.5 mM) or high (28 mM) glucose concentrations to stimulate C-peptide secretion and the response was compared with human pancreatic islets (islets). Human C-peptide secretion in hBTSCs in KM was not affected by high glucose stimulation (data are means ± SD of 6 experiments). In hBTSCs cultured for 14 days in KM containing 0.1 μM Pdx1, C-peptide secretion was detected at low glucose levels and further increased after exposure to high glucose concentration (data are means ± SD of 6 experiments; *p< 0.05). Human pancreatic islets (islets) were used as a positive control of the high glucose challenge (data are means ± SD of 6 experiments; *p< 0.05).</p>", "links"=>[], "tags"=>["hepatocyte cell line", "Adult Human Biliary Tree Stem Cells Differentiate", "Pdx 1 exposure", "hBTSC", "Pdx 1 protein", "pdx", "Pdx 1", "HepG 2 cell line", "time polymerase chain reaction", "pancreatic islet cells", "Pdx 1 peptide", "Recombinant Human Pdx 1 Peptide Generation"], "article_id"=>1505893, "categories"=>["Biological Sciences"], "users"=>["Vincenzo Cardinale", "Rosa Puca", "Guido Carpino", "Gaia Scafetta", "Anastasia Renzi", "Michele De Canio", "Francesca Sicilia", "Lorenzo Nevi", "Domenico Casa", "Rocco Panetta", "Pasquale Bartolomeo Berloco", "Lola M. Reid", "Giorgio Federici", "Eugenio Gaudio", "Marella Maroder", "Domenico Alvaro"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0134677.g004", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Pdx1_induced_hBTSC_derived_946_pancreatic_islets_are_functioning_active_/1505893", "title"=>"Pdx1 induced hBTSC-derived β-pancreatic islets are functioning active.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-08-07 03:32:39"}

PMC Usage Stats | Further Information

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Relative Metric

{"start_date"=>"2015-01-01T00:00:00Z", "end_date"=>"2015-12-31T00:00:00Z", "subject_areas"=>[]}

F1000Prime