Attenuation of Hind-Limb Ischemia in Mice with Endothelial-Like Cells Derived from Different Sources of Human Stem Cells
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{"title"=>"Attenuation of Hind-Limb Ischemia in Mice with Endothelial-Like Cells Derived from Different Sources of Human Stem Cells", "type"=>"journal", "authors"=>[{"first_name"=>"Wing-Hon", "last_name"=>"Lai"}, {"first_name"=>"Jenny C. Y.", "last_name"=>"Ho"}, {"first_name"=>"Yau-Chi", "last_name"=>"Chan"}, {"first_name"=>"Joyce H. L.", "last_name"=>"Ng"}, {"first_name"=>"Ka-Wing", "last_name"=>"Au"}, {"first_name"=>"Lai-Yung", "last_name"=>"Wong"}, {"first_name"=>"Chung-Wah", "last_name"=>"Siu"}, {"first_name"=>"Hung-Fat", "last_name"=>"Tse"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"doi"=>"10.1371/journal.pone.0057876", "pmid"=>"23472116", "issn"=>"1932-6203", "isbn"=>"1932-6203"}, "id"=>"abf4af1b-e96b-326b-8f8c-fc2bbb9c3fb6", "abstract"=>"Functional endothelial-like cells (EC) have been successfully derived from different cell sources and potentially used for treatment of cardiovascular diseases; however, their relative therapeutic efficacy remains unclear. We differentiated functional EC from human bone marrow mononuclear cells (BM-EC), human embryonic stem cells (hESC-EC) and human induced pluripotent stem cells (hiPSC-EC), and compared their in-vitro tube formation, migration and cytokine expression profiles, and in-vivo capacity to attenuate hind-limb ischemia in mice. Successful differentiation of BM-EC was only achieved in 1/6 patient with severe coronary artery disease. Nevertheless, BM-EC, hESC-EC and hiPSC-EC exhibited typical cobblestone morphology, had the ability of uptaking DiI-labeled acetylated low-density-lipoprotein, and binding of Ulex europaeus lectin. In-vitro functional assay demonstrated that hiPSC-EC and hESC-EC had similar capacity for tube formation and migration as human umbilical cord endothelial cells (HUVEC) and BM-EC (P>0.05). While increased expression of major angiogenic factors including epidermal growth factor, hepatocyte growth factor, vascular endothelial growth factor, placental growth factor and stromal derived factor-1 were observed in all EC cultures during hypoxia compared with normoxia (P<0.05), the magnitudes of cytokine up-regulation upon hypoxic were more dramatic in hiPSC-EC and hESC-EC (P<0.05). Compared with medium, transplanting BM-EC (n = 6), HUVEC (n = 6), hESC-EC (n = 8) or hiPSC-EC (n = 8) significantly attenuated severe hind-limb ischemia in mice via enhancement of neovascularization. In conclusion, functional EC can be generated from hECS and hiPSC with similar therapeutic efficacy for attenuation of severe hind-limb ischemia. Differentiation of functional BM-EC was more difficult to achieve in patients with cardiovascular diseases, and hESC-EC or iPSC-EC are readily available as \"off-the-shelf\" format for the treatment of tissue ischemia.", "link"=>"http://www.mendeley.com/research/attenuation-hindlimb-ischemia-mice-endotheliallike-cells-derived-different-sources-human-stem-cells", "reader_count"=>47, "reader_count_by_academic_status"=>{"Researcher"=>8, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>11, "Student > Master"=>7, "Other"=>7, "Student > Bachelor"=>8, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1, "Professor"=>2}, "reader_count_by_user_role"=>{"Researcher"=>8, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>11, "Student > Master"=>7, "Other"=>7, "Student > Bachelor"=>8, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1, "Professor"=>2}, "reader_count_by_subject_area"=>{"Engineering"=>4, "Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>10, "Medicine and Dentistry"=>16, "Agricultural and Biological Sciences"=>15, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>4}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>16}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>15}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>10}, "Unspecified"=>{"Unspecified"=>1}}, "reader_count_by_country"=>{"United States"=>2}, "group_count"=>13}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/977360"], "description"=>"<p>(<b>A</b>) Laser Doppler scanning was preformed immediately after ligation of femoral artery to confirm the induction of hind-limb ischemia. Then repeated Doppler scanning was performed at Day 7, 14, 21 and 28 after transplantation of 3 million hESC-EC, two different hiPSC-ECs, BM-EC, HUVEC or medium alone into the ischemic muscle. Representative photos from the laser Doppler scanning at Day 0, Day 14 and Day 28 were shown. Blue color represents lower perfusion and red color represents higher perfusion (upper panel). In each animal, limb perfusion was quantified and the average of the perfusion ratio against the control limb was calculated in different time point (lower panel). As compared with medium injection, there was progressive and significant improvement of blood flow over the ischemic limbs after hESC-EC, hiPSC-ECs, BM-EC and HUVEC transplantation (n = 6 in each group, *p<0.05 vs. medium control at different time points). However, there were no significant differences in the blood flow over the ischemic limbs among those mice after hESC-EC, hiPSC-EC and BM-EC transplantation (p>0.05 at different time points). (<b>B</b>) Representative photos show the immunohistochemical staining for von Willebrand factor (vWF) and smooth muscle actin (SMA) of histological sections over the ischemic limbs at Day 28 after hESC-EC, hiPSC-EC, BM-EC or medium injection (upper panel). The number of vessels with positive staining (brown color) for vWF and SMA were determined under a light microscope with ×400 magnification with 20 random fields. As compared with medium injection, there were significant higher number of vessels with vWF positive but not SMA over the ischemic limbs after hESC-EC, hiPSC-ECs and BM-EC transplantation (*p vs. medium control and * vs HUVEC, p<0.05). However, there were no significant differences in the number of vessels with vWF and SMA positive over the ischemic limbs among those mice after hESC-EC, hiPSC-EC and BM-EC transplantation (p>0.05).</p>", "links"=>[], "tags"=>["perfusion", "ischemic", "hind-limbs", "transplantation", "derived", "endothelial-like"], "article_id"=>644640, "categories"=>["Cell Biology", "Developmental Biology"], "users"=>["Wing-Hon Lai", "Jenny C. Y. Ho", "Yau-Chi Chan", "Joyce H. L. Ng", "Ka-Wing Au", "Lai-Yung Wong", "Chung-Wah Siu", "Hung-Fat Tse"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0057876.g004", "stats"=>{"downloads"=>3, "page_views"=>28, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Improvement_of_blood_perfusion_in_ischemic_hind_limbs_after_transplantation_of_derived_endothelial_like_cells_/644640", "title"=>"Improvement of blood perfusion in ischemic hind-limbs after transplantation of derived endothelial-like cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-06 08:17:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/977349"], "description"=>"<p>(A) Tube formation assay was performed with 1×10<sup>4</sup> cells seeded onto Matrigel and the presence of micro vessel formation was visualized by phase contrast microscope (100×) after 6 hours of incubation at 37°C. There was no significant difference in length of tube-like vessels formed by hESC-EC, hiPSC-EC, BM-EC and HUVEC (p>0.05). (<b>B</b>) Modified Boyden chamber assay was used to determine the migration ability of the hESC-EC, hiPSC-EC, BM-EC and HUVEC. These cells were placed in the inner chamber and allowed to migrate across the porous membrane toward the outer chamber with the presence of medium alone (control) or medium supplemented with vascular endothelial growth factor (VEGF). Then, the numbers of migrated cells per mm<sup>2</sup> present in the transwell were counted under light microscope (100×, upper panel). The numbers of migrated cells were significantly increased after supplement with VEGF in hESC-EC, hiPSC-EC, BM-EC and HUVEC as compared with their corresponding control (*p<0.05). However, there were no differences among those EC in the number of migrated cells with the presence of medium alone or medium supplemented with VEGF (p<0.05, lower panel).</p>", "links"=>[], "tags"=>["derived", "endothelial-like"], "article_id"=>644634, "categories"=>["Cell Biology", "Developmental Biology"], "users"=>["Wing-Hon Lai", "Jenny C. Y. Ho", "Yau-Chi Chan", "Joyce H. L. Ng", "Ka-Wing Au", "Lai-Yung Wong", "Chung-Wah Siu", "Hung-Fat Tse"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0057876.g002", "stats"=>{"downloads"=>1, "page_views"=>37, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_vitro_functional_assessment_of_derived_endothelial_like_cells_/644634", "title"=>"<i>In- vitro</i> functional assessment of derived endothelial-like cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-06 08:15:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/1011554"], "description"=>"<p>CAD = coronary arteries diseases; CHF = chronic heart failure; HT = hypertension; DM = diabetes mellitus; Lipid = hyperlipidemia; PCI = percutaneous coronary intervention; CABG = coronary artery bypass graft, ‘+’ = present, ‘−’ = absent.</p>", "links"=>[], "tags"=>["marrow", "mnc"], "article_id"=>671962, "categories"=>["Cell Biology", "Developmental Biology"], "users"=>["Wing-Hon Lai", "Jenny C. Y. Ho", "Yau-Chi Chan", "Joyce H. L. Ng", "Ka-Wing Au", "Lai-Yung Wong", "Chung-Wah Siu", "Hung-Fat Tse"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0057876.t001", "stats"=>{"downloads"=>6, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Clinical_characteristic_of_the_patients_8217_marrow_MNC_used_in_this_study_/671962", "title"=>"Clinical characteristic of the patients’ marrow MNC used in this study.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-03-05 00:32:42"}
  • {"files"=>["https://ndownloader.figshare.com/files/977376"], "description"=>"<p>(<b>A</b>) Representative photos show the immunohistochemical staining for human nuclear antigen (HNA) over the ischemic hind-limb at Day 7 and 28 after hESC-EC, hiPSC-1-EC, BM-EC or medium injection (upper panel). Red arrow indicated engrafted hESC-EC, hiPSC-EC, BM-EC in the ischemic limb with HNA positive (brown), indicating those cells were from human origin. At Day 7, clusters of HNA positive cells were observed at the injection sites after hESC-EC, hiPSC-1-EC, and BM-EC transplantation but not after medium injection. At Day 28, only a few HNA positive cells were found around the vasculature at the injection sites after hESC-EC, hiPSC-1-EC, and BM-EC transplantation. As compared with Day 28, the numbers of HNA positive cells detected at the injection sites were significantly higher at Day 7 after hESC-EC, hiPSC-1-EC, BM-EC transplantation (*p<0.05, lower panel). (<b>B</b>) Representative photos showing live cells tracking by quantifying luciferase activity using the Xenogen’s <i>in vivo</i> optical imaging systems at Day 7, 14 and 28 after hESC-EC, hiPSC-1-EC, or medium injection (upper panel). Luciferase activity was plot as total flux measurement in photons/second/cm<sup>2</sup>. Declined in intensity of luciferase activity over time after hESC-EC and hiPSC-1-EC transplantation was shown, suggesting progressive cell loss (n = 4 in each group, lower panel).</p>", "links"=>[], "tags"=>["derived", "endothelial-like", "cells", "14", "28"], "article_id"=>644646, "categories"=>["Cell Biology", "Developmental Biology"], "users"=>["Wing-Hon Lai", "Jenny C. Y. Ho", "Yau-Chi Chan", "Joyce H. L. Ng", "Ka-Wing Au", "Lai-Yung Wong", "Chung-Wah Siu", "Hung-Fat Tse"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0057876.g005", "stats"=>{"downloads"=>1, "page_views"=>31, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Retention_of_derived_endothelial_like_cells_at_Day_7_14_and_28_after_transplantation_/644646", "title"=>"Retention of derived endothelial-like cells at Day 7, 14 and 28 after transplantation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-06 08:18:38"}
  • {"files"=>["https://ndownloader.figshare.com/files/977354"], "description"=>"<p>A total of 1×10<sup>5</sup> cells were plated into 6-well gelatin coated plate for overnight incubation, then the cells were washed and cultivated in EBM2 basal medium without supplement for 5 days under normoxic (20% O<sub>2</sub>) or hypoxic (0.1% O<sub>2</sub>). Cytokine profiling of their conditioned mediums obtained were determined by Procarta multiplex bead-based immunoassay kit. (A) Bar charts and (B) multivariate radar chart showed the expression of epidermal growth factor (EGF), basic fibroblast growth factor (FGF-B), hepatocyte growth factor (HGF), leptin, vascular endothelial growth factor (VEGF), placental growth factor (PIGF) and stromal derived factor-1 (SDF-1); adhesion molecules including, ICAM1 and vascular cell adhesion molecule (VCAM) in the conditioned medium upon normoxic and hypoxic treatment.</p>", "links"=>[], "tags"=>["cytokine", "profiles", "conditioned", "derived", "endothelial-like", "cells", "normoxic", "hypoxic"], "article_id"=>644636, "categories"=>["Cell Biology", "Developmental Biology"], "users"=>["Wing-Hon Lai", "Jenny C. Y. Ho", "Yau-Chi Chan", "Joyce H. L. Ng", "Ka-Wing Au", "Lai-Yung Wong", "Chung-Wah Siu", "Hung-Fat Tse"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0057876.g003", "stats"=>{"downloads"=>1, "page_views"=>39, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Angiogenic_cytokine_profiles_of_conditioned_medium_obtained_from_derived_endothelial_like_cells_under_normoxic_and_hypoxic_condition_/644636", "title"=>"Angiogenic cytokine profiles of conditioned medium obtained from derived endothelial-like cells under normoxic and hypoxic condition.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-06 08:16:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/977344"], "description"=>"<p>(A) Phase contrast photos (left panel) showing the morphology of hESC-EC, hiPSC-EC, bone marrow derived EC (BM-EC) and human umbilical cord endothelial cell (HUVEC) in a confluent culture. Immunofluorescence staining showed the expression of typical endothelial cells markers including <i>Ulex europaeus</i> lectin (green) and uptake of Di-Acetyl-LDL (red) (second panel) in hESC-EC, hiPSC-1-EC, BM-EC and HUVEC. Furthermore, expression of CD31, von Willebrand factor (vWF) and intercellular adhesion molecule 1 (ICAM) were also detected in hESC-EC, hiPSC-EC, BM-EC and HUVEC (B) Flow cytometry analysis revealed the expression of CD31, vWF, Kinase insert domain receptor (KDR) in hESC-EC, hiPSC-EC, BM-EC and HUVEC (left panel). There were no differences in the percentage of cells with expression of CD31, vWF, KDR and co-expression of CD31 and KDR between hESC-EC, hiPSC-EC and BM-EC (right panel).</p>", "links"=>[], "tags"=>["derived", "endothelial-like", "cells"], "article_id"=>644630, "categories"=>["Cell Biology", "Developmental Biology"], "users"=>["Wing-Hon Lai", "Jenny C. Y. Ho", "Yau-Chi Chan", "Joyce H. L. Ng", "Ka-Wing Au", "Lai-Yung Wong", "Chung-Wah Siu", "Hung-Fat Tse"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0057876.g001", "stats"=>{"downloads"=>1, "page_views"=>23, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Characterization_of_derived_endothelial_like_cells_EC_/644630", "title"=>"Characterization of derived endothelial-like cells (EC).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-06 08:13:58"}

PMC Usage Stats | Further Information

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Relative Metric

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