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2 protocols using ter119 pe cyanine 7

1

Identification of Cardiac Mesenchymal Stem Cells

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Expression of surface markers or green fluorescent protein (GFP) in the cultured CD51+cMSCs was tested using Influx flow cytometers (BD, United States). Assessment of a CD51-positive subpopulation with other subpopulations (Sca-1/C-kit/CD31/CD45/Ter19) in the heart tissue was also analyzed using flow cytometry. Data were analyzed with FlowJo 7.5 software (Treestar, United States). The following anti-mouse antibodies were used: CD51-PE (BD), Ter119-PE Cyanine 7 (eBioscience), CD44-PE (eBioscience), CD106-Alexa Fluor 647 (eBioscience), CD90 (BD Biosciences), Sca-1-APC (eBioscience), C-kit-APC-eFluor 780 (eBioscience), CD34-FITC (eBioscience), and CD11b-APC (eBioscience). The antibodies were used at the dilutions recommended by the manufacturers.
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2

Flow Cytometric Analyses of Immune Cells

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Flow cytometric analyses were performed with Influx or Gallios (Beckman Coulter) flow cytometers. The data were analysed with FlowJo v10 software (TreeStar) or CytExpert2.3 software (Beckman Coulter). The following anti‐mouse antibodies were used: CD44‐PE (IM7), CD73‐APC (AD2), CD90‐PE (30‐H12), CD106‐Alexa Fluor 647 (429), CD45‐PE‐Cyanine7 (30‐F11), CD11b‐PE‐Cyanine7 (M1/70), PDGFRα‐APC (APA5), IL‐17A‐PE‐Cyanine7 (eBio17B7), TER‐119‐PE‐Cyanine7 (TER‐119), and IL‐22‐PE (1H8PWSR) from eBioscience; CD3‐V450 (500A2), CD8‐BV510 (53–6.7), CD31‐APC (MEC 13.3), CD34‐BV421 (RAM34), CD105‐BV421 (MJ7/18) Sca‐1‐V450 (D7), TNF‐α‐PE (MP6‐XT22) and IFN‐γ‐APC (XMG1.2) from BD; and CD4‐APCCY7 (GK1.5) from BioLegend.
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