One-Step Derivation of Mesenchymal Stem Cell (MSC)-Like Cells from Human Pluripotent Stem Cells on a Fibrillar Collagen Coating
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{"title"=>"One-step derivation of mesenchymal stem cell (MSC)-like cells from human pluripotent stem cells on a fibrillar collagen coating", "type"=>"journal", "authors"=>[{"first_name"=>"Yongxing", "last_name"=>"Liu", "scopus_author_id"=>"26661198600"}, {"first_name"=>"A. Jon", "last_name"=>"Goldberg", "scopus_author_id"=>"7402481507"}, {"first_name"=>"James E.", "last_name"=>"Dennis", "scopus_author_id"=>"7202979992"}, {"first_name"=>"Gloria A.", "last_name"=>"Gronowicz", "scopus_author_id"=>"7003450103"}, {"first_name"=>"Liisa T.", "last_name"=>"Kuhn", "scopus_author_id"=>"26637035200"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"364480380", "sgr"=>"84863347182", "issn"=>"19326203", "pmid"=>"22457746", "scopus"=>"2-s2.0-84863347182", "doi"=>"10.1371/journal.pone.0033225", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)"}, "id"=>"4b5736c3-017a-3926-a549-38d1b05e5341", "abstract"=>"Controlled differentiation of human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) into cells that resemble adult mesenchymal stem cells (MSCs) is an attractive approach to obtain a readily available source of progenitor cells for tissue engineering. The present study reports a new method to rapidly derive MSC-like cells from hESCs and hiPSCs, in one step, based on culturing the cells on thin, fibrillar, type I collagen coatings that mimic the structure of physiological collagen. Human H9 ESCs and HDFa-YK26 iPSCs were singly dissociated in the presence of ROCK inhibitor Y-27632, plated onto fibrillar collagen coated plates and cultured in alpha minimum essential medium (alpha-MEM) supplemented with 10% fetal bovine serum, 50 uM magnesium L-ascorbic acid phosphate and 100 nM dexamethasone. While fewer cells attached on the collagen surface initially than standard tissue culture plastic, after culturing for 10 days, resilient colonies of homogenous spindle-shaped cells were obtained. Flow cytometric analysis showed that a high percentage of the derived cells expressed typical MSC surface markers including CD73, CD90, CD105, CD146 and CD166 and were negative as expected for hematopoietic markers CD34 and CD45. The MSC-like cells derived from pluripotent cells were successfully differentiated in vitro into three different lineages: osteogenic, chondrogenic, and adipogenic. Both H9 hES and YK26 iPS cells displayed similar morphological changes during the derivation process and yielded MSC-like cells with similar properties. In conclusion, this study demonstrates that bioimimetic, fibrillar, type I collagen coatings applied to cell culture plates can be used to guide a rapid, efficient derivation of MSC-like cells from both human ES and iPS cells.", "link"=>"http://www.mendeley.com/research/onestep-derivation-mesenchymal-stem-cell-msclike-cells-human-pluripotent-stem-cells-fibrillar-collag", "reader_count"=>86, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>7, "Researcher"=>17, "Student > Doctoral Student"=>11, "Student > Ph. D. Student"=>29, "Student > Postgraduate"=>2, "Student > Master"=>10, "Other"=>2, "Student > Bachelor"=>2, "Lecturer"=>2, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>7, "Researcher"=>17, "Student > Doctoral Student"=>11, "Student > Ph. D. Student"=>29, "Student > Postgraduate"=>2, "Student > Master"=>10, "Other"=>2, "Student > Bachelor"=>2, "Lecturer"=>2, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>4, "Agricultural and Biological Sciences"=>37, "Veterinary Science and Veterinary Medicine"=>1, "Chemistry"=>1, "Computer Science"=>1, "Economics, Econometrics and Finance"=>2, "Engineering"=>7, "Biochemistry, Genetics and Molecular Biology"=>9, "Materials Science"=>1, "Medicine and Dentistry"=>20, "Neuroscience"=>1, "Psychology"=>1, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Materials Science"=>{"Materials Science"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>20}, "Psychology"=>{"Psychology"=>1}, "Unspecified"=>{"Unspecified"=>4}, "Engineering"=>{"Engineering"=>7}, "Chemistry"=>{"Chemistry"=>1}, "Neuroscience"=>{"Neuroscience"=>1}, "Economics, Econometrics and Finance"=>{"Economics, Econometrics and Finance"=>2}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>37}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>9}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"Canada"=>1, "Iran"=>1, "United States"=>1, "Japan"=>1, "Germany"=>1, "Russia"=>1}, "group_count"=>1}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/664820"], "description"=>"<p>Surface antigen profile analysis by flow cytometry for the derived MSC-like cells. (A) and (C) are from cells derived from H9 at passage 2 and 4, respectively. (B) and (D) are from cells derived from YK26-iPSCs at passage 2 and 4, respectively. The black histogram represents the unstained control and white overlay represents each antigen. The numeric values shown within the histograms represent the percentage of the positive cells.</p>", "links"=>[], "tags"=>["cytometry", "markers"], "article_id"=>335309, "categories"=>["Biotechnology", "Cell Biology", "Developmental Biology"], "users"=>["Yongxing Liu", "A. Jon Goldberg", "James E. Dennis", "Gloria A. Gronowicz", "Liisa T. Kuhn"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0033225.g003", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Flow_cytometry_analysis_of_surface_markers_consistent_with_adult_MSCs_/335309", "title"=>"Flow cytometry analysis of surface markers consistent with adult MSCs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 07:29:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/664561"], "description"=>"<p>Scanning electron microscopy of the bioimimetic fibrillar type I collagen coating shown prior to cell seeding (scale bar: 5 µm).</p>", "links"=>[], "tags"=>["collagen", "substrate", "leads", "fibrillar"], "article_id"=>335037, "categories"=>["Biotechnology", "Cell Biology", "Developmental Biology"], "users"=>["Yongxing Liu", "A. Jon Goldberg", "James E. Dennis", "Gloria A. Gronowicz", "Liisa T. Kuhn"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0033225.g001", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Self_assembly_of_Type_I_collagen_on_a_tissue_culture_plastic_substrate_leads_to_a_thin_fibrillar_coating_/335037", "title"=>"Self-assembly of Type I collagen on a tissue culture plastic substrate leads to a thin fibrillar coating.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 07:27:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/665170"], "description"=>"<p>Oil red O staining of the 21 day adipogenic cultures was positive in both (A) MSC-like cells derived from H9-hESC and (B) MSC-like cells derived from YK26-iPS cells. Scale bars: 0.1 mm. (C) RT-qPCR analysis for mRNA expression of adipogenic genes PPARγ and FABP4. 1: undifferentiated H9-hESC; 2 and 3: MSC-like cells that derived from H9-hESC and YK26-iPS cells, respectively, before adipogenic differentiation; 4 and 5: 21 day adipogenic cultures of the MSC-like cells derived from H9-hESC and YK26-iPS cells, respectively.</p>", "links"=>[], "tags"=>["adipogenic"], "article_id"=>335656, "categories"=>["Biotechnology", "Cell Biology", "Developmental Biology"], "users"=>["Yongxing Liu", "A. Jon Goldberg", "James E. Dennis", "Gloria A. Gronowicz", "Liisa T. Kuhn"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0033225.g006", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_vitro_adipogenic_differentiation_/335656", "title"=>"<i>In vitro</i> adipogenic differentiation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 07:31:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/665053"], "description"=>"<p>Positive chondrogenic matrix staining observed after chondrogenic differentiation. (A) and (B) are images of 21d cultures of MSC-like cells derived from H9-hESC and YK26-iPSCs, respectively. Alcian blue staining of the histological sections of the chondrogenic pellet cultures was positive. Immunohistochemical staining for aggrecan and collagen Type II on the histological sections of the 21 d chondrogenic pellet cultures was positive. Scale bars: 0.2 mm. (C) Negative control for Col II. (D) RT-qPCR analysis for mRNA expression of chondrogenic genes including SOX9, COL2A1 and aggrecan (ACAN). 1: undifferentiated H9-hESC; 2 and 3: MSC-like cells that derived from H9-hESC and YK26-iPS cells, respectively, before chondrogenic differentiation; 4 and 5: 21 day chondrogenic cultures of the MSC-like cells derived from H9-hESC and YK26-iPS cells, respectively.</p>", "links"=>[], "tags"=>["chondrogenic"], "article_id"=>335541, "categories"=>["Biotechnology", "Cell Biology", "Developmental Biology"], "users"=>["Yongxing Liu", "A. Jon Goldberg", "James E. Dennis", "Gloria A. Gronowicz", "Liisa T. Kuhn"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0033225.g005", "stats"=>{"downloads"=>3, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_vitro_chondrogenic_differentiation_/335541", "title"=>"<i>In vitro</i> chondrogenic differentiation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 07:30:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/664932"], "description"=>"<p>Phase contrast images of the cells in the osteogenic cultures on day 2 and 35. (A) and (B) show MSC-like cells derived from H9-hESC and YK26-iPSCs, respectively. The 35 day cultures were stained with xylenol orange dye (XO) to reveal mineral deposition. The red fluorescence in the last column of images displays the positive XO staining. Scale bars: 0.2 mm. (C) RT-qPCR analysis for the mRNA expression of osteogenic-related genes including RUNX2, COL1A1 and ALP on day 35. 1: undifferentiated H9-hESC; 2 and 3: MSC-like cells that derived from H9-hESC and YK26-iPS cells, respectively, before osteogenic differentiation; 4 and 5: 35 day osteogenic cultures of the MSC-like cells derived from H9-hESC and YK26-iPS cells, respectively.</p>", "links"=>[], "tags"=>["osteogenic"], "article_id"=>335421, "categories"=>["Biotechnology", "Cell Biology", "Developmental Biology"], "users"=>["Yongxing Liu", "A. Jon Goldberg", "James E. Dennis", "Gloria A. Gronowicz", "Liisa T. Kuhn"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0033225.g004", "stats"=>{"downloads"=>2, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_vitro_osteogenic_differentiation_/335421", "title"=>"<i>In vitro</i> osteogenic differentiation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 07:29:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/664677"], "description"=>"<p>Phase contrast images of cell morphology as a function of cell culture plate coating chemistry. Note that all the images were taken from H9-hESC cultures, but are also representative for the corresponding YK26-iPSCs cultures. (A) The typical fibroblast-like morphology of cells obtained on the collagen coating after 10 d (P0). The cells are located in separate colonies and some areas of the plate remain open. (B) The morphology of the highly confluent cells obtained on tissue culture treated polystyrene after 10 days. The inset shows an enlarged region (2×) of the image; arrowheads indicate the tight junction-like structure formed between cells. (C) The clear fibroblast-like morphology of cells subcultured (P1) on collagen coating for 4 days. (D) The varied morphology of cells derived for 10 days on either tissue culture treated polystyrene or collagen coatings after plating cell colonies rather than single cells; the arrow indicates the protrusion growing up into the medium. Scale bars for phase contrast images: 0.2 mm.</p>", "links"=>[], "tags"=>["morphologies", "observed"], "article_id"=>335159, "categories"=>["Biotechnology", "Cell Biology", "Developmental Biology"], "users"=>["Yongxing Liu", "A. Jon Goldberg", "James E. Dennis", "Gloria A. Gronowicz", "Liisa T. Kuhn"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0033225.g002", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Various_cell_morphologies_were_observed_in_response_to_the_cell_culture_plate_coatings_/335159", "title"=>"Various cell morphologies were observed in response to the cell culture plate coatings.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 07:28:37"}

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Relative Metric

{"start_date"=>"2012-01-01T00:00:00Z", "end_date"=>"2012-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences", "average_usage"=>[322, 550, 671, 773, 864, 955, 1048, 1135, 1223, 1308, 1387, 1465, 1534, 1602, 1673, 1744, 1813, 1885, 1955, 2026, 2093, 2160, 2228, 2290, 2349]}, {"subject_area"=>"/Biology and life sciences/Cell biology", "average_usage"=>[319, 556, 679, 785, 881, 970, 1062, 1149, 1236, 1323, 1402, 1474, 1545, 1617, 1681, 1754, 1822, 1892, 1963, 2031, 2099, 2165, 2233, 2299, 2359]}, {"subject_area"=>"/Engineering and technology", "average_usage"=>[308, 500, 607, 700, 782, 876, 957, 1039, 1128, 1214, 1290, 1365, 1431, 1496, 1560, 1634, 1695, 1772, 1839, 1908, 1972, 2043, 2106, 2179, 2246]}]}
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