Perivascular-Like Cells Contribute to the Stability of the Vascular Network of Osteogenic Tissue Formed from Cell Sheet-Based Constructs
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{"title"=>"Perivascular-like cells contribute to the stability of the vascular network of osteogenic tissue formed from cell sheet-based constructs", "type"=>"journal", "authors"=>[{"first_name"=>"Luís F.", "last_name"=>"Mendes", "scopus_author_id"=>"55317145400"}, {"first_name"=>"Rogério P.", "last_name"=>"Pirraco", "scopus_author_id"=>"24473477700"}, {"first_name"=>"Wojciech", "last_name"=>"Szymczyk", "scopus_author_id"=>"55317676400"}, {"first_name"=>"Ana M.", "last_name"=>"Frias", "scopus_author_id"=>"26646425800"}, {"first_name"=>"Tírcia C.", "last_name"=>"Santos", "scopus_author_id"=>"22935620100"}, {"first_name"=>"Rui L.", "last_name"=>"Reis", "scopus_author_id"=>"56861715700"}, {"first_name"=>"Alexandra P.", "last_name"=>"Marques", "scopus_author_id"=>"22035647000"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84864053203", "doi"=>"10.1371/journal.pone.0041051", "pui"=>"365284986", "pmid"=>"22829909", "scopus"=>"2-s2.0-84864053203", "issn"=>"19326203", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)"}, "id"=>"300fc754-ed53-3067-bec6-bbe76a776983", "abstract"=>"In recent years several studies have been supporting the existence of a close relationship in terms of function and progeny between Mesenchymal Stem Cells (MSCs) and Pericytes. This concept has opened new perspectives for the application of MSCs in Tissue Engineering (TE), with special interest for the pre-vascularization of cell dense constructs. In this work, cell sheet technology was used to create a scaffold-free construct composed of osteogenic, endothelial and perivascular-like (CD146(+)) cells for improved in vivo vessel formation, maturation and stability. The CD146 pericyte-associated phenotype was induced from human bone marrow mesenchymal stem cells (hBMSCs) by the supplementation of standard culture medium with TGF-β1. Co-cultured cell sheets were obtained by culturing perivascular-like (CD146(+)) cells and human umbilical vein endothelial cells (HUVECs) on an hBMSCs monolayer maintained in osteogenic medium for 7 days. The perivascular-like (CD146(+)) cells and the HUVECs migrated and organized over the collagen-rich osteogenic cell sheet, suggesting the existence of cross-talk involving the co-cultured cell types. Furthermore the presence of that particular ECM produced by the osteoblastic cells was shown to be the key regulator for the singular observed organization. The osteogenic and angiogenic character of the proposed constructs was assessed in vivo. Immunohistochemistry analysis of the explants revealed the integration of HUVECs with the host vasculature as well as the osteogenic potential of the created construct, by the expression of osteocalcin. Additionally, the analysis of the diameter of human CD146 positive blood vessels showed a higher mean vessel diameter for the co-cultured cell sheet condition, reinforcing the advantage of the proposed model regarding blood vessels maturation and stability and for the in vitro pre-vascularization of TE constructs.", "link"=>"http://www.mendeley.com/research/perivascularlike-cells-contribute-stability-vascular-network-osteogenic-tissue-formed-cell-sheetbase", "reader_count"=>54, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Researcher"=>11, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>20, "Student > Postgraduate"=>2, "Student > Master"=>7, "Other"=>2, "Student > Bachelor"=>6, "Lecturer > Senior Lecturer"=>1, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Researcher"=>11, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>20, "Student > Postgraduate"=>2, "Student > Master"=>7, "Other"=>2, "Student > Bachelor"=>6, "Lecturer > Senior Lecturer"=>1, "Professor"=>2}, "reader_count_by_subject_area"=>{"Engineering"=>10, "Unspecified"=>3, "Biochemistry, Genetics and Molecular Biology"=>3, "Materials Science"=>3, "Agricultural and Biological Sciences"=>19, "Medicine and Dentistry"=>12, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Physics and Astronomy"=>1, "Chemistry"=>1, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>10}, "Materials Science"=>{"Materials Science"=>3}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>12}, "Chemistry"=>{"Chemistry"=>1}, "Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>19}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>3}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"Iran"=>1, "United Kingdom"=>2, "Australia"=>1}, "group_count"=>2}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/606811"], "description"=>"<p>Single osteogenic cell sheet derived from hBMSCs A) after H&E staining and immunostained for (B) osteocalcin and (C) type-I collagen; Co-cultured cell sheets after (E) H&E staining and immunostaining for (F) CD31, (G) CD146, (H) osteocalcin and (I) Type-I collagen. Identification of positive signal was determined in comparison to immunocytochemistry negative controls (D;J). * PVDF membrane used to protect cell sheet during processing.</p>", "links"=>[], "tags"=>["characterization", "co-cultured", "sheets", "14", "days", "osteogenic", "detachment"], "article_id"=>277293, "categories"=>["Biotechnology", "Cell Biology", "Developmental Biology"], "users"=>["Luís F. Mendes", "Rogério P. Pirraco", "Wojciech Szymczyk", "Ana M. Frias", "Tírcia C. Santos", "Rui L. Reis", "Alexandra P. Marques"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0041051.g003", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Histological_characterization_of_single_and_co_cultured_cell_sheets_after_14_days_in_culture_in_osteogenic_medium_and_after_detachment_by_temperature_decrease_and_contraction_/277293", "title"=>"Histological characterization of single and co-cultured cell sheets after 14 days in culture in osteogenic medium and after detachment by temperature decrease and contraction.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-07-19 02:01:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/606729"], "description"=>"<p>hBMSCs were seeded and cultured for 7 days in osteogenic medium in thermoresponsive dishes. To obtain co-cultured CS, HUVECs and perivascular-like (CD146<sup>+</sup>) cells were cultured, at a ratio of 4∶1, on the osteogenic CS in M199 supplemented with osteogenic factors for further 7 days (experimental). Control homotypic osteogenic CS were maintained in osteogenic medium. At day 14, CS were retrieved from the thermoresponsive dishes by temperature decrease and the experimental model was built by stacking of a homotypic osteogenic CS onto the co-cultured CS using a poly(vinylidene difluoride) (PVDF) membrane.</p>", "links"=>[], "tags"=>["methodology", "stacked", "co-cultured", "sheets"], "article_id"=>277214, "categories"=>["Biotechnology", "Cell Biology", "Developmental Biology"], "users"=>["Luís F. Mendes", "Rogério P. Pirraco", "Wojciech Szymczyk", "Ana M. Frias", "Tírcia C. Santos", "Rui L. Reis", "Alexandra P. Marques"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0041051.g002", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_vitro_culture_methodology_to_obtain_a_stacked_co_cultured_cell_sheets_CS_based_model_/277214", "title"=>"<i>In vitro</i> culture methodology to obtain a stacked co-cultured cell sheets (CS)-based model.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-07-19 02:00:14"}
  • {"files"=>["https://ndownloader.figshare.com/files/606614"], "description"=>"<p>(A) CD146 expression of bone marrow mononuclear fraction at isolation day; (B) CD146 expression on hBMSCs (P5) cultured in complete α-MEM; (C; D) CD146 expression analysis, by flow cytometry (C) and immunocytochemistry (green) (D), on hBMSCs (P5) cultured in complete α-MEM supplemented with 1 ng/mL TGF-β1 for 7 days; Evolution of cell morphology of hBMSCs (E) before and (F) after culture in α-MEM +1 ng/mL TGF-β1 for 7 days. For immunocytochemistry DAPI (blue) was used as nuclear staining. Right upper corner image in D represent a higher magnification.</p>", "links"=>[], "tags"=>["cytometry", "immunocytochemistry", "marrow", "derived", "cells", "passages", "cultured"], "article_id"=>277098, "categories"=>["Biotechnology", "Cell Biology", "Developmental Biology"], "users"=>["Luís F. Mendes", "Rogério P. Pirraco", "Wojciech Szymczyk", "Ana M. Frias", "Tírcia C. Santos", "Rui L. Reis", "Alexandra P. Marques"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0041051.g001", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Representative_flow_cytometry_and_immunocytochemistry_analysis_of_human_bone_marrow_derived_cells_at_different_passages_and_cultured_with_and_without_TGF_946_1_/277098", "title"=>"Representative flow cytometry and immunocytochemistry analysis of human bone marrow derived cells at different passages and cultured with and without TGF-β1.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-07-19 01:58:18"}
  • {"files"=>["https://ndownloader.figshare.com/files/607155"], "description"=>"<p>(A–D) H&E staining on (A;B )control and (C;D) experimental explants after 7 (A;C) and 21 days (B;D) of subcutaneous implantation showing their localization and morphology. (E–L) Immunolocalization of osteocalcin on (E;F) control and (G;H) experimental explants at 7 (E;G) and 21 (F;H) days of implantation revealing osteogenic commitment on both test conditions. (I–L) immunostaining negative control of respective E–H conditions.</p>", "links"=>[], "tags"=>["staining", "osteocalcin", "immunolocalization", "explants", "21", "days", "transplantation", "sheet-based"], "article_id"=>277638, "categories"=>["Biotechnology", "Cell Biology", "Developmental Biology"], "users"=>["Luís F. Mendes", "Rogério P. Pirraco", "Wojciech Szymczyk", "Ana M. Frias", "Tírcia C. Santos", "Rui L. Reis", "Alexandra P. Marques"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0041051.g005", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_H_amp_E_staining_and_osteocalcin_immunolocalization_on_explants_retrieved_7_and_21_days_after_transplantation_of_cell_sheet_based_constructs_/277638", "title"=>"H&E staining and osteocalcin immunolocalization on explants retrieved 7 and 21 days after transplantation of cell sheet-based constructs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-07-19 02:07:18"}
  • {"files"=>["https://ndownloader.figshare.com/files/316794", "https://ndownloader.figshare.com/files/316859", "https://ndownloader.figshare.com/files/316894"], "description"=>"<div><p>In recent years several studies have been supporting the existence of a close relationship in terms of function and progeny between Mesenchymal Stem Cells (MSCs) and Pericytes. This concept has opened new perspectives for the application of MSCs in Tissue Engineering (TE), with special interest for the pre-vascularization of cell dense constructs. In this work, cell sheet technology was used to create a scaffold-free construct composed of osteogenic, endothelial and perivascular-like (CD146<sup>+</sup>) cells for improved in vivo vessel formation, maturation and stability. The CD146 pericyte-associated phenotype was induced from human bone marrow mesenchymal stem cells (hBMSCs) by the supplementation of standard culture medium with TGF-β1. Co-cultured cell sheets were obtained by culturing perivascular-like (CD146<sup>+</sup>) cells and human umbilical vein endothelial cells (HUVECs) on an hBMSCs monolayer maintained in osteogenic medium for 7 days. The perivascular-like (CD146<sup>+</sup>) cells and the HUVECs migrated and organized over the collagen-rich osteogenic cell sheet, suggesting the existence of cross-talk involving the co-cultured cell types. Furthermore the presence of that particular ECM produced by the osteoblastic cells was shown to be the key regulator for the singular observed organization. The osteogenic and angiogenic character of the proposed constructs was assessed <em>in vivo</em>. Immunohistochemistry analysis of the explants revealed the integration of HUVECs with the host vasculature as well as the osteogenic potential of the created construct, by the expression of osteocalcin. Additionally, the analysis of the diameter of human CD146 positive blood vessels showed a higher mean vessel diameter for the co-cultured cell sheet condition, reinforcing the advantage of the proposed model regarding blood vessels maturation and stability and for the <em>in vitro</em> pre-vascularization of TE constructs.</p> </div>", "links"=>[], "tags"=>["perivascular-like", "cells", "vascular", "osteogenic", "formed", "sheet-based", "constructs"], "article_id"=>122524, "categories"=>["Biotechnology", "Cell Biology", "Developmental Biology"], "users"=>["Luís F. Mendes", "Rogério P. Pirraco", "Wojciech Szymczyk", "Ana M. Frias", "Tírcia C. Santos", "Rui L. Reis", "Alexandra P. Marques"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0041051.s001", "https://dx.doi.org/10.1371/journal.pone.0041051.s002", "https://dx.doi.org/10.1371/journal.pone.0041051.s003"], "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Perivascular_Like_Cells_Contribute_to_the_Stability_of_the_Vascular_Network_of_Osteogenic_Tissue_Formed_from_Cell_Sheet_Based_Constructs/122524", "title"=>"Perivascular-Like Cells Contribute to the Stability of the Vascular Network of Osteogenic Tissue Formed from Cell Sheet-Based Constructs", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-07-19 00:42:04"}
  • {"files"=>["https://ndownloader.figshare.com/files/606973"], "description"=>"<p>(A,B) Co-cultures on hBMSCs-derived osteogenic cells showing endothelial colonies (red) and elongated perivascular-like (CD146<sup>+</sup>) cells (green) interacting with HUVECs and with them-self (Arrow). (C) Co-cultures of HUVECs (red) and perivascular-like (CD146<sup>+</sup>) cells (green) on plastic adherent conditions showing random organization. (D) Confluent layer of hBMSCs-derived osteogenic cells lacking the expression of CD146. DAPI (blue) was used as nuclear staining.</p>", "links"=>[], "tags"=>["cd146", "dil-acldl", "uptake", "hbmscs", "monocultures", "co-cultures", "perivascular-like", "cells", "14", "days"], "article_id"=>277450, "categories"=>["Biotechnology", "Cell Biology", "Developmental Biology"], "users"=>["Luís F. Mendes", "Rogério P. Pirraco", "Wojciech Szymczyk", "Ana M. Frias", "Tírcia C. Santos", "Rui L. Reis", "Alexandra P. Marques"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0041051.g004", "stats"=>{"downloads"=>2, "page_views"=>161, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunocytochemistry_for_CD146_expression_and_Dil_AcLDL_uptake_by_hBMSCs_monocultures_and_co_cultures_with_perivascular_like_CD146_cells_and_HUVECs_after_14_days_of_culture_/277450", "title"=>"Immunocytochemistry for CD146 expression and Dil-AcLDL uptake by hBMSCs monocultures and co-cultures with perivascular-like (CD146<sup>+</sup>) cells and HUVECs, after 14 days of culture.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-07-19 02:04:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/607330"], "description"=>"<p>Immunohistochemistry for (A;B) CD31 and (C–F) CD146 on (C;D) control and (E,F) experimental conditions at days 7 (C;E) and 21 (D;F) of implantation; (G–J) Immunostaining negative control of respective conditions. → negative blood vessels for CD146; ▸ positive blood vessel for CD146. (K) Human cells (green) detected using human-specific anti-mitochondria antibodies on the experimental condition 7 days after implantation. (L) Co-localization (yellow) of CD146 (red) and human-specific anti-mitochondria (green) revealed cellular assembling in a blood vessel-like structure (arrow) on the experimental condition 7 days after implantation. DAPI (blue) was used as nuclear staining. Representation of (M) the mean diameter and (N) the number of CD146 positive vessels present on control and experimental conditions at days 7 and 21 of implantation. *p≤0.05; **p≤0.01.</p>", "links"=>[], "tags"=>["transplanted", "sheet-based"], "article_id"=>277820, "categories"=>["Biotechnology", "Cell Biology", "Developmental Biology"], "users"=>["Luís F. Mendes", "Rogério P. Pirraco", "Wojciech Szymczyk", "Ana M. Frias", "Tírcia C. Santos", "Rui L. Reis", "Alexandra P. Marques"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0041051.g006", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Angiogenic_potential_of_the_transplanted_cell_sheet_based_constructs_/277820", "title"=>"Angiogenic potential of the transplanted cell sheet-based constructs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-07-19 02:10:20"}

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Relative Metric

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