Assessing the Permeability of Engineered Capillary Networks in a 3D Culture
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Mendeley | Further Information

{"title"=>"Assessing the permeability of engineered capillary networks in a 3D culture", "type"=>"journal", "authors"=>[{"first_name"=>"Stephanie J.", "last_name"=>"Grainger", "scopus_author_id"=>"57196016844"}, {"first_name"=>"Andrew J.", "last_name"=>"Putnam", "scopus_author_id"=>"7005981221"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"21760956", "sgr"=>"79960077120", "doi"=>"10.1371/journal.pone.0022086", "scopus"=>"2-s2.0-79960077120", "pui"=>"362097301", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "issn"=>"19326203"}, "id"=>"ac3a517b-6967-35e0-9476-2b545e24248c", "abstract"=>"Many pathologies are characterized by poor blood vessel growth and reduced nutrient delivery to the surrounding tissue, introducing a need for tissue engineered blood vessels. Our lab has developed a 3D co-culture method to grow interconnected networks of pericyte-invested capillaries, which can anastamose with host vasculature following implantation to restore blood flow to ischemic tissues. However, if the engineered vessels contain endothelial cells (ECs) that are misaligned or contain wide junctional gaps, they may function improperly and behave more like the pathologic vessels that nourish tumors. The purpose of this study was to test the resistance to permeability of these networks in vitro, grown with different stromal cell types, as a metric of vessel functionality. A fluorescent dextran tracer was used to visualize transport across the endothelium and the pixel intensity was quantified using a customized MATLAB algorithm. In fibroblast-EC co-cultures, the dextran tracer easily penetrated through the vessel wall and permeability was high through the first 5 days of culture, indicative of vessel immaturity. Beyond day 5, dextran accumulated at the periphery of the vessel, with very little transported across the endothelium. Quantitatively, permeability dropped from initial levels of 61% to 39% after 7 days, and to 7% after 2 weeks. When ECs were co-cultured with bone marrow-derived mesenchymal stem cells (MSCs) or adipose-derived stem cells (AdSCs), much tighter control of permeability was achieved. Relative to the EC-fibroblast co-cultures, permeabilities were reduced 41% for the EC-MSC co-cultures and 50% for the EC-AdSC co-cultures after 3 days of culture. By day 14, these permeabilities decreased by 68% and 77% over the EC-fibroblast cultures. Co-cultures containing stem cells exhibit elevated VE-cadherin levels and more prominent EC-EC junctional complexes when compared to cultures containing fibroblasts. These data suggest the stromal cell identity influences the functionality and physiologic relevance of engineered capillary networks.", "link"=>"http://www.mendeley.com/research/assessing-permeability-engineered-capillary-networks-3d-culture", "reader_count"=>75, "reader_count_by_academic_status"=>{"Unspecified"=>3, "Professor > Associate Professor"=>8, "Researcher"=>12, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>27, "Student > Postgraduate"=>1, "Student > Master"=>7, "Other"=>3, "Student > Bachelor"=>7, "Lecturer"=>1, "Professor"=>3}, "reader_count_by_user_role"=>{"Unspecified"=>3, "Professor > Associate Professor"=>8, "Researcher"=>12, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>27, "Student > Postgraduate"=>1, "Student > Master"=>7, "Other"=>3, "Student > Bachelor"=>7, "Lecturer"=>1, "Professor"=>3}, "reader_count_by_subject_area"=>{"Engineering"=>22, "Unspecified"=>6, "Biochemistry, Genetics and Molecular Biology"=>3, "Materials Science"=>4, "Agricultural and Biological Sciences"=>29, "Medicine and Dentistry"=>9, "Chemistry"=>2}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>22}, "Materials Science"=>{"Materials Science"=>4}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>9}, "Chemistry"=>{"Chemistry"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>29}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>6}}, "reader_count_by_country"=>{"United States"=>5, "Denmark"=>1, "Brazil"=>1, "Germany"=>4}, "group_count"=>4}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/759209"], "description"=>"<p>Quantification generated via the customized MATLAB algorithm to determine the amount of Texas Red-conjugated dextran within the capillary interior at each time point. An isolated * symbol indicates statistical significance (p<0.05) between data points (NHLF vs. MSC vs. AdSC) at any one particular time point, whereas a bar beneath the * symbol indicates statistical significance (p<0.05) across all 3 experimental groups at each time point. The +, #, and % symbols indicate statistically significant differences (p<0.005) between the data marked by these symbols at certain time points and all other time points marked with different symbols.</p>", "links"=>[], "tags"=>["permeability", "customized"], "article_id"=>429590, "categories"=>["Physiology", "Biotechnology"], "users"=>["Stephanie J. Grainger", "Andrew J. Putnam"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022086.g005", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Quantification_of_permeability_using_a_customized_algorithm_/429590", "title"=>"Quantification of permeability using a customized algorithm.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:58:44"}
  • {"files"=>["https://ndownloader.figshare.com/files/759004"], "description"=>"<p>60x confocal microscopy images of days 5 (A) and 10 (B) capillaries after 70 kDa dextran tracer addition (red), fixation with 10% formalin, and human CD31 staining (green) of HUVECs. Inset images: HUVECs (top) and Dextran (bottom) show hollow lumens with or without dextran. A) Red tracer is present within the lumens of the capillaries, demonstrating a low resistance to permeability. B) Red tracer is excluded from the lumens of the capillaries, demonstrating an increased resistance to permeability as the capillaries mature over time.</p>", "links"=>[], "tags"=>["tracer", "localization", "confocal"], "article_id"=>429385, "categories"=>["Physiology", "Biotechnology"], "users"=>["Stephanie J. Grainger", "Andrew J. Putnam"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022086.g003", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Dextran_tracer_localization_via_confocal_microscopy_/429385", "title"=>"Dextran tracer localization via confocal microscopy.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:57:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/759407"], "description"=>"<p>A–C) Representative 4x images of fluorescently-tagged HUVECs forming capillary networks in co-culture with A) AdSCs, B) MSCs, C) NHLFs. D) Comparison of growth kinetics for several stromal cell types in the bead assay. “Mono” denotes the stromal cells were cultured as a monolayer on top of the fibrin gel, a fixed distance away from the HUVEC-coated beads; “distributed” indicates the stromal cells were distributed throughout the entire 3D gel. E) Comparison of the number of vessel segments per network for each stromal cell type. The * symbol indicates statistical significance (p<0.05) between time points within a given culture condition. The +, #, %, and @ symbols indicate statistical significance (p<0.005) for a particular culture condition at a given time point versus the other culture conditions. For example, the total lengths of the capillary networks induced by distributed NHLFs are significantly different than those induced by the monolayer NHLFs and those induced by all of the other cell types and distributions (denoted by #).</p>", "links"=>[], "tags"=>["kinetics", "stromal"], "article_id"=>429786, "categories"=>["Physiology", "Biotechnology"], "users"=>["Stephanie J. Grainger", "Andrew J. Putnam"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022086.g007", "stats"=>{"downloads"=>4, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Capillary_growth_kinetics_vary_by_stromal_cell_type_/429786", "title"=>"Capillary growth kinetics vary by stromal cell type.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:59:50"}
  • {"files"=>["https://ndownloader.figshare.com/files/758879"], "description"=>"<p>In Panel A, the algorithm makes a 3D overlay of the images of each channel. The user defines the location of a background box. In Panel B, each green image from the 3D z-stack is depicted in gray scale. In Panel C, the maximum projection of the green z-stack is converted to a binary image after the background (defined in Panel A) is eliminated. In Panel D, the portion of the red 3D z-stack that corresponds to the same area depicted in the green image is shown in gray scale as a maximum projection. In Panel E, only those pixels from the red image in Panel D that overlap with the green pixels in Panel C are shown. The pixel intensities from the image in Panel E are tallied and output as a ratio of red:green.</p>", "links"=>[], "tags"=>["biotechnology", "physiology"], "article_id"=>429259, "categories"=>["Physiology", "Biotechnology"], "users"=>["Stephanie J. Grainger", "Andrew J. Putnam"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022086.g002", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Pixel_comparison_algorithm_/429259", "title"=>"Pixel comparison algorithm.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:56:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/759278"], "description"=>"<p>A–C) 20x confocal microscopy images of day 14 capillaries from co-cultures of EC-NHLF (A), EC-AdSC (B), and EC-MSC (C) after staining for VE-cadherin (green) and DAPI (blue). D) Western blot showing expression of VE-cadherin within each of the co-culture conditions.</p>", "links"=>[], "tags"=>["stromal"], "article_id"=>429660, "categories"=>["Physiology", "Biotechnology"], "users"=>["Stephanie J. Grainger", "Andrew J. Putnam"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022086.g006", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_VE_cadherin_organization_and_expression_vary_as_a_function_of_stromal_cell_type_/429660", "title"=>"VE-cadherin organization and expression vary as a function of stromal cell type.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:59:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/758794"], "description"=>"<p>Texas Red-conjugated dextran (70 kDa) is added to the bulk gel for 30 minutes and allowed to freely diffuse. Early capillary sprouts, which lack mature cell-cell junctions between the endothelial cells that form the tubules, will easily allow the tracer to be transported from the tissue space into the interior of the capillary. By contrast, mature capillaries that are stabilized by pericytes and possess more mature cell-cell junctions are capable of excluding the tracer from the lumen.</p>", "links"=>[], "tags"=>["inverse", "permeability", "capillary", "functionality", "kinetics", "3d"], "article_id"=>429165, "categories"=>["Physiology", "Biotechnology"], "users"=>["Stephanie J. Grainger", "Andrew J. Putnam"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022086.g001", "stats"=>{"downloads"=>2, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Model_of_inverse_permeability_used_to_determine_capillary_functionality_kinetics_in_a_3D_culture_/429165", "title"=>"Model of inverse permeability used to determine capillary functionality kinetics in a 3D culture.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:56:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/759125"], "description"=>"<p>Histamine treatment quantitatively modulates permeability of the engineered capillary networks, as expected. In this graph, the * symbol indicates statistical significance (p<0.05) between the black and white bars (−/+ histamine) at each time point. The +, #, and % symbols indicate statistically significant differences (p<0.005) between the data marked by these symbols at certain time points and all other time points marked with different symbols. For example, the values at day 3 and day 5 (for both −/+ histamine) are significantly different from the values at all of the other time points, but not from each other.</p>", "links"=>[], "tags"=>["validation", "histamine", "modulate", "capillary"], "article_id"=>429503, "categories"=>["Physiology", "Biotechnology"], "users"=>["Stephanie J. Grainger", "Andrew J. Putnam"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0022086.g004", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Assay_validation_using_histamine_to_modulate_capillary_permeability_/429503", "title"=>"Assay validation using histamine to modulate capillary permeability.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:58:17"}

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Relative Metric

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