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{"title"=>"Natural scaffolds for renal differentiation of human embryonic stem cells for kidney tissue engineering", "type"=>"journal", "authors"=>[{"first_name"=>"Cynthia A.", "last_name"=>"Batchelder", "scopus_author_id"=>"11739745100"}, {"first_name"=>"Michele L.", "last_name"=>"Martinez", "scopus_author_id"=>"35105403200"}, {"first_name"=>"Alice F.", "last_name"=>"Tarantal", "scopus_author_id"=>"7005209142"}], "year"=>2015, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-84955469309", "sgr"=>"84955469309", "pui"=>"607909909", "pmid"=>"26645109", "doi"=>"10.1371/journal.pone.0143849"}, "id"=>"f3083fe8-3920-38dd-a1f3-4e874707f8d1", "abstract"=>"Despite the enthusiasm for bioengineering of functional renal tissues for transplantation, many obstacles remain before the potential of this technology can be realized in a clinical setting. Viable tissue engineering strategies for the kidney require identification of the necessary cell populations, efficient scaffolds, and the 3D culture conditions to develop and support the unique architecture and physiological function of this vital organ. Our studies have previously demonstrated that decellularized sections of rhesus monkey kidneys of all age groups provide a natural extracellular matrix (ECM) with sufficient structural properties with spatial and organizational influences on human embryonic stem cell (hESC) migration and differentiation. To further explore the use of decellularized natural kidney scaffolds for renal tissue engineering, pluripotent hESC were seeded in whole- or on sections of kidney ECM and cell migration and phenotype compared with the established differentiation assays for hESC. Results of qPCR and immunohistochemical analyses demonstrated upregulation of renal lineage markers when hESC were cultured in decellularized scaffolds without cytokine or growth factor stimulation, suggesting a role for the ECM in directing renal lineage differentiation. hESC were also differentiated with growth factors and compared when seeded on renal ECM or a new biologically inert polysaccharide scaffold for further maturation. Renal lineage markers were progressively upregulated over time on both scaffolds and hESC were shown to express signature genes of renal progenitor, proximal tubule, endothelial, and collecting duct populations. These findings suggest that natural scaffolds enhance expression of renal lineage markers particularly when compared to embryoid body culture. The results of these studies show the capabilities of a novel polysaccharide scaffold to aid in defining a protocol for renal progenitor differentiation from hESC, and advance the promise of tissue engineering as a source of functional kidney tissue.", "link"=>"http://www.mendeley.com/research/natural-scaffolds-renal-differentiation-human-embryonic-stem-cells-kidney-tissue-engineering-3", "reader_count"=>48, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>2, "Researcher"=>6, "Student > Ph. D. Student"=>7, "Student > Postgraduate"=>1, "Student > Master"=>12, "Other"=>2, "Student > Bachelor"=>13, "Professor"=>3}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>2, "Researcher"=>6, "Student > Ph. D. Student"=>7, "Student > Postgraduate"=>1, "Student > Master"=>12, "Other"=>2, "Student > Bachelor"=>13, "Professor"=>3}, "reader_count_by_subject_area"=>{"Unspecified"=>4, "Engineering"=>5, "Biochemistry, Genetics and Molecular Biology"=>8, "Materials Science"=>2, "Medicine and Dentistry"=>6, "Agricultural and Biological Sciences"=>15, "Pharmacology, Toxicology and Pharmaceutical Science"=>2, "Chemical Engineering"=>2, "Physics and Astronomy"=>1, "Chemistry"=>2, "Social Sciences"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>5}, "Materials Science"=>{"Materials Science"=>2}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>6}, "Chemistry"=>{"Chemistry"=>2}, "Social Sciences"=>{"Social Sciences"=>1}, "Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>15}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>8}, "Unspecified"=>{"Unspecified"=>4}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>2}, "Chemical Engineering"=>{"Chemical Engineering"=>2}}, "reader_count_by_country"=>{"Germany"=>1}, "group_count"=>2}

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• http://doi.org/10.1021/acsbiomaterials.7b00083
• http://doi.org/10.1038/s41390-019-0736-6
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• {"id"=>"674485191344279552", "text"=>"#PLOSONE: Natural Scaffolds for Renal Differentiation of Human Embryonic Stem Cells for Kidney Tissue Engineering https://t.co/vmCqX0Hpnt", "created_at"=>"2015-12-09T07:06:15Z", "user"=>"PLOSSynbio", "user_name"=>"PLOS Synbio", "user_profile_image"=>"http://pbs.twimg.com/profile_images/508712140245454848/RTAKdRsW_normal.jpeg"}
• {"id"=>"674508314068299776", "text"=>"RT @PLOSSynbio: #PLOSONE: Natural Scaffolds for Renal Differentiation of Human Embryonic Stem Cells for Kidney Tissue Engineering https://t…", "created_at"=>"2015-12-09T08:38:08Z", "user"=>"JCoutoPhoenix", "user_name"=>"Jillian Couto", "user_profile_image"=>"http://pbs.twimg.com/profile_images/563372531671760896/Y_e9JI06_normal.jpeg"}

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Figshare

• {"files"=>["https://ndownloader.figshare.com/files/4299385"], "description"=>"<p><b>(A-D).</b> H&E staining of cell-scaffold constructs after 20 days in culture with two growth factor protocols (A or B). (<b>E-H</b>). Tubule-like structures were present and positive for CK but not VIM. (<b>I-L).</b> Regions positive for PAX2 (intermediate mesoderm and induced metanephric mesenchyme) were typically distinct from regions positive for WT1 (induced mesenchyme) under Protocol A. These markers were expressed in a more diffuse pattern throughout the construct under Protocol B. Tubule-like structures in Protocol B with renal ECM were WT1-positive and surrounded by a ring of PAX2-positive cells. (<b>M-P</b>). The proximal tubule marker AQP1 was expressed with greater frequency under Protocol A and on PSS while the Loop of Henle marker UMOD was rarely expressed in any construct. (<b>Q-T</b>). The distal tubule/collecting duct marker ECAD was widely expressed on cells with epithelial morphology in all constructs while the ascending Loop of Henle and collecting duct marker CALB was only expressed in constructs under Protocol A with PSS. (<b>U-X</b>). The proximal tubule marker EMA was expressed on some tubule structures in all constructs with small vessel-like structures positive for the endothelial marker CD31. Nuclei were visualized with DAPI (blue). Representative images shown; N≥3 experiments. Scale bar = 100 μm.</p>", "links"=>[], "tags"=>["tissue engineering", "differentiation", "lineage markers", "growth factor stimulation", "Viable tissue engineering strategies", "decellularized", "hESC", "ECM", "novel polysaccharide scaffold", "Kidney Tissue Engineering", "3 D culture conditions", "Human Embryonic Stem Cells", "Renal lineage markers", "embryoid body culture", "rhesus monkey kidneys"], "article_id"=>1621857, "categories"=>["Biophysics", "Space Science", "Medicine", "Cell Biology", "Genetics", "Biotechnology", "Ecology", "Biological Sciences not elsewhere classified", "Developmental Biology", "Marine Biology"], "users"=>["Cynthia A. Batchelder", "Michele L. Martinez", "Alice F. Tarantal"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0143849.g005"], "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Renal_directed_differentiation_of_hESC_on_renal_ECM_or_PSS_/1621857", "title"=>"Renal-directed differentiation of hESC on renal ECM or PSS.", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2015-12-08 07:42:52"}
• {"files"=>["https://ndownloader.figshare.com/files/4299388"], "description"=>"<p>Pluripotent hESC were plated in suspension cultures to form embryoid bodies with supplemental growth factors as shown. Gene expression (qPCR) relative to the housekeeping gene EF1α was calculated using undifferentiated day 0 cells as the comparator. After 12 days of suspension culture, embryoid bodies were collected and plated on renal ECM or PSS at the air-medium interface in basal medium (arrows). Both scaffolds were equal or superior to embryoid body culture in upregulating renal lineage genes (N≥3 replicates). With the exception of SIX2, PSS were equal (OSR1) or superior (BRACHY, LIM1, WT1, PAX2) to renal ECM in supporting renal precursor populations.</p>", "links"=>[], "tags"=>["tissue engineering", "differentiation", "lineage markers", "growth factor stimulation", "Viable tissue engineering strategies", "decellularized", "hESC", "ECM", "novel polysaccharide scaffold", "Kidney Tissue Engineering", "3 D culture conditions", "Human Embryonic Stem Cells", "Renal lineage markers", "embryoid body culture", "rhesus monkey kidneys"], "article_id"=>1621859, "categories"=>["Biophysics", "Space Science", "Medicine", "Cell Biology", "Genetics", "Biotechnology", "Ecology", "Biological Sciences not elsewhere classified", "Developmental Biology", "Marine Biology"], "users"=>["Cynthia A. Batchelder", "Michele L. Martinez", "Alice F. Tarantal"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0143849.g006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_of_early_renal_lineage_markers_in_differentiating_hESC_cultured_under_Protocol_A_/1621859", "title"=>"Expression of early renal lineage markers in differentiating hESC cultured under Protocol A.", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2015-12-08 07:42:52"}
• {"files"=>["https://ndownloader.figshare.com/files/4299394"], "description"=>"<p>Pluripotent hESC were differentiated in suspension cultures as embryoid bodies with supplemental growth factors as shown. After 5 days, cells were plated on renal ECM or PSS at the air-medium interface in basal medium (arrows). Gene expression (qPCR) relative to the housekeeping gene EF1α was calculated using undifferentiated day 0 cells as the comparator (N≥3 replicates). Both types of scaffolds were superior to embryoid body cultures in directing upregulation of intermediate mesoderm (OSR1, LIM1, PAX2) and the metanephric mesenchyme gene, WT1. Renal ECM was similar to embryoid body culture in expression of BRY and SIX2, while these genes were upregulated on PSS.</p>", "links"=>[], "tags"=>["tissue engineering", "differentiation", "lineage markers", "growth factor stimulation", "Viable tissue engineering strategies", "decellularized", "hESC", "ECM", "novel polysaccharide scaffold", "Kidney Tissue Engineering", "3 D culture conditions", "Human Embryonic Stem Cells", "Renal lineage markers", "embryoid body culture", "rhesus monkey kidneys"], "article_id"=>1621860, "categories"=>["Biophysics", "Space Science", "Medicine", "Cell Biology", "Genetics", "Biotechnology", "Ecology", "Biological Sciences not elsewhere classified", "Developmental Biology", "Marine Biology"], "users"=>["Cynthia A. Batchelder", "Michele L. Martinez", "Alice F. Tarantal"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0143849.g007"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_of_early_renal_lineage_markers_in_differentiating_hESC_cultured_under_Protocol_B_/1621860", "title"=>"Expression of early renal lineage markers in differentiating hESC cultured under Protocol B.", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2015-12-08 07:42:52"}
• {"files"=>["https://ndownloader.figshare.com/files/4299355", "https://ndownloader.figshare.com/files/4299358"], "description"=>"<div><p>Despite the enthusiasm for bioengineering of functional renal tissues for transplantation, many obstacles remain before the potential of this technology can be realized in a clinical setting. Viable tissue engineering strategies for the kidney require identification of the necessary cell populations, efficient scaffolds, and the 3D culture conditions to develop and support the unique architecture and physiological function of this vital organ. Our studies have previously demonstrated that decellularized sections of rhesus monkey kidneys of all age groups provide a natural extracellular matrix (ECM) with sufficient structural properties with spatial and organizational influences on human embryonic stem cell (hESC) migration and differentiation. To further explore the use of decellularized natural kidney scaffolds for renal tissue engineering, pluripotent hESC were seeded in whole- or on sections of kidney ECM and cell migration and phenotype compared with the established differentiation assays for hESC. Results of qPCR and immunohistochemical analyses demonstrated upregulation of renal lineage markers when hESC were cultured in decellularized scaffolds without cytokine or growth factor stimulation, suggesting a role for the ECM in directing renal lineage differentiation. hESC were also differentiated with growth factors and compared when seeded on renal ECM or a new biologically inert polysaccharide scaffold for further maturation. Renal lineage markers were progressively upregulated over time on both scaffolds and hESC were shown to express signature genes of renal progenitor, proximal tubule, endothelial, and collecting duct populations. These findings suggest that natural scaffolds enhance expression of renal lineage markers particularly when compared to embryoid body culture. The results of these studies show the capabilities of a novel polysaccharide scaffold to aid in defining a protocol for renal progenitor differentiation from hESC, and advance the promise of tissue engineering as a source of functional kidney tissue.</p></div>", "links"=>[], "tags"=>["tissue engineering", "differentiation", "lineage markers", "growth factor stimulation", "Viable tissue engineering strategies", "decellularized", "hESC", "ECM", "novel polysaccharide scaffold", "Kidney Tissue Engineering", "3 D culture conditions", "Human Embryonic Stem Cells", "Renal lineage markers", "embryoid body culture", "rhesus monkey kidneys"], "article_id"=>1621861, "categories"=>["Biophysics", "Space Science", "Medicine", "Cell Biology", "Genetics", "Biotechnology", "Ecology", "Biological Sciences not elsewhere classified", "Developmental Biology", "Marine Biology"], "users"=>["Cynthia A. Batchelder", "Michele L. Martinez", "Alice F. Tarantal"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0143849.s001", "https://dx.doi.org/10.1371/journal.pone.0143849.s002"], "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Natural_Scaffolds_for_Renal_Differentiation_of_Human_Embryonic_Stem_Cells_for_Kidney_Tissue_Engineering_/1621861", "title"=>"Natural Scaffolds for Renal Differentiation of Human Embryonic Stem Cells for Kidney Tissue Engineering", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2015-12-08 07:42:52"}
• {"files"=>["https://ndownloader.figshare.com/files/4299367"], "description"=>"<p>Whole-kidney scaffolds before (<b>A</b>) and after (<b>B</b>) perfusion with 1% SDS. Kidneys from the same donor were compared after whole-kidney perfusion (<b>C</b>) or static section (<b>D</b>) decellularization methods. Histological comparison of decellularized whole kidney (<b>E</b>) with native kidney (<b>F</b>) demonstrated removal of cellular contents with maintenance of native renal architecture. The section decellularization process at 4°C without agitation (<b>G</b>) was greatly improved when sections were placed on a shaker during the decellularization process at room temperature (<b>H</b>). Scale bars = 100 μm.</p>", "links"=>[], "tags"=>["tissue engineering", "differentiation", "lineage markers", "growth factor stimulation", "Viable tissue engineering strategies", "decellularized", "hESC", "ECM", "novel polysaccharide scaffold", "Kidney Tissue Engineering", "3 D culture conditions", "Human Embryonic Stem Cells", "Renal lineage markers", "embryoid body culture", "rhesus monkey kidneys"], "article_id"=>1621838, "categories"=>["Biophysics", "Space Science", "Medicine", "Cell Biology", "Genetics", "Biotechnology", "Ecology", "Biological Sciences not elsewhere classified", "Developmental Biology", "Marine Biology"], "users"=>["Cynthia A. Batchelder", "Michele L. Martinez", "Alice F. Tarantal"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0143849.g001"], "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Production_of_decellularized_renal_scaffolds_/1621838", "title"=>"Production of decellularized renal scaffolds.", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2015-12-08 07:42:52"}
• {"files"=>["https://ndownloader.figshare.com/files/4299373"], "description"=>"<p>(<b>A</b>) Schematic of custom-designed, perfusion bioreactor for cell seeding via the renal artery (RA) or ureter (U). (<b>B</b>) Recellularized kidneys after 7 days of culture. Cells seeded via the renal artery (<b>C-D</b>) or ureter (<b>E-F</b>) were observed in medullary vascular or tubular lumens but not in outer cortical tubules or glomeruli. With longer culture periods enhanced recellularization was observed (<b>G-H</b>), particularly in medullary regions. Representative images from a minimum of three independent experiments are shown. Medulla (M), Cortex (Cx), scale bars = 100 μm.</p>", "links"=>[], "tags"=>["tissue engineering", "differentiation", "lineage markers", "growth factor stimulation", "Viable tissue engineering strategies", "decellularized", "hESC", "ECM", "novel polysaccharide scaffold", "Kidney Tissue Engineering", "3 D culture conditions", "Human Embryonic Stem Cells", "Renal lineage markers", "embryoid body culture", "rhesus monkey kidneys"], "article_id"=>1621840, "categories"=>["Biophysics", "Space Science", "Medicine", "Cell Biology", "Genetics", "Biotechnology", "Ecology", "Biological Sciences not elsewhere classified", "Developmental Biology", "Marine Biology"], "users"=>["Cynthia A. Batchelder", "Michele L. Martinez", "Alice F. Tarantal"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0143849.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Recellularization_of_kidney_scaffolds_with_hESC_/1621840", "title"=>"Recellularization of kidney scaffolds with hESC.", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2015-12-08 07:42:52"}
• {"files"=>["https://ndownloader.figshare.com/files/4299400"], "description"=>"<p>Antibodies for immunohistochemical analysis.</p>", "links"=>[], "tags"=>["tissue engineering", "differentiation", "lineage markers", "growth factor stimulation", "Viable tissue engineering strategies", "decellularized", "hESC", "ECM", "novel polysaccharide scaffold", "Kidney Tissue Engineering", "3 D culture conditions", "Human Embryonic Stem Cells", "Renal lineage markers", "embryoid body culture", "rhesus monkey kidneys"], "article_id"=>2645716, "categories"=>["Biophysics", "Space Science", "Medicine", "Cell Biology", "Genetics", "Biotechnology", "Ecology", "Biological Sciences not elsewhere classified", "Developmental Biology", "Marine Biology"], "users"=>["Cynthia A. Batchelder", "Michele L. Martinez", "Alice F. Tarantal"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0143849.t001", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Antibodies_for_immunohistochemical_analysis_/2645716", "title"=>"Antibodies for immunohistochemical analysis.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2015-12-08 07:42:52"}
• {"files"=>["https://ndownloader.figshare.com/files/4299376"], "description"=>"<p>hESC were differentiated as embryoid bodies (<b>A</b>) or cultured in whole kidneys (<b>B</b>) or sections of kidneys (<b>C</b>) where cells were typically observed in the medulla and medullary rays. (<b>D-F</b>) Renal developmental markers WT1 and PAX2 were upregulated in whole or sections of decellularized kidneys when compared with embryoid body differentiation. (<b>G-I</b>) AQP1, a marker of proximal tubules, was expressed in tubule-like structures in embryoid bodies and kidney sections, but not in whole kidneys. (<b>J-L</b>) Vimentin, a mesenchymal and mesangial marker, was expressed under all culture conditions. Other markers of mature renal cell types including SMA (mesangial and vascular smooth muscle marker) and Calbindin (renal distal tubules) were not expressed. Nuclei were visualized with DAPI (blue); scale bars = 100 μm.</p>", "links"=>[], "tags"=>["tissue engineering", "differentiation", "lineage markers", "growth factor stimulation", "Viable tissue engineering strategies", "decellularized", "hESC", "ECM", "novel polysaccharide scaffold", "Kidney Tissue Engineering", "3 D culture conditions", "Human Embryonic Stem Cells", "Renal lineage markers", "embryoid body culture", "rhesus monkey kidneys"], "article_id"=>1621850, "categories"=>["Biophysics", "Space Science", "Medicine", "Cell Biology", "Genetics", "Biotechnology", "Ecology", "Biological Sciences not elsewhere classified", "Developmental Biology", "Marine Biology"], "users"=>["Cynthia A. Batchelder", "Michele L. Martinez", "Alice F. Tarantal"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0143849.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Renal_developmental_markers_are_upregulated_by_renal_ECM_/1621850", "title"=>"Renal developmental markers are upregulated by renal ECM.", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2015-12-08 07:42:52"}
• {"files"=>["https://ndownloader.figshare.com/files/4299379"], "description"=>"<p>Relative expression of mesodermal (BRY), posterior primitive streak (OSR1), intermediate mesodermal (PAX2), and metanephric mesenchymal (WT1) genes in hESC differentiated as embryoid bodies or in renal scaffolds. Mean ± standard error of the mean (SEM); N≥3 replicates.</p>", "links"=>[], "tags"=>["tissue engineering", "differentiation", "lineage markers", "growth factor stimulation", "Viable tissue engineering strategies", "decellularized", "hESC", "ECM", "novel polysaccharide scaffold", "Kidney Tissue Engineering", "3 D culture conditions", "Human Embryonic Stem Cells", "Renal lineage markers", "embryoid body culture", "rhesus monkey kidneys"], "article_id"=>1621852, "categories"=>["Biophysics", "Space Science", "Medicine", "Cell Biology", "Genetics", "Biotechnology", "Ecology", "Biological Sciences not elsewhere classified", "Developmental Biology", "Marine Biology"], "users"=>["Cynthia A. Batchelder", "Michele L. Martinez", "Alice F. Tarantal"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0143849.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Early_renal_lineage_genes_were_upregulated_by_renal_ECM_/1621852", "title"=>"Early renal lineage genes were upregulated by renal ECM.", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2015-12-08 07:42:52"}

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