Standard flow cytometry was performed to determine the presence of MSC markers, as defined by the International Society for Cellular Therapy.15 UC‐MSCs were stained using a Human MSC Analysis kit (BD Biosciences, Franklin Lakes, NJ, USA) containing the following mouse monoclonal antibodies, which are positive markers for MSCs: fluorescein isothiocyanate (FITC)‐conjugated anti‐human CD90; phycoerythrin (PE)‐conjugated anti‐human CD105; allophycocyanin (APC)‐conjugated anti‐human CD73; FITC‐conjugated anti‐human CD44 (BD Biosciences), and PE‐conjugated anti‐HLA‐ABC (BD Biosciences). Additionally, UC‐MSCs were stained with FITC‐conjugated anti‐HLA‐DR (BD Biosciences), FITC‐conjugated anti‐human CD34 (BD Biosciences), PE‐conjugated anti‐human CD11b (BD Biosciences), PE‐conjugated anti‐human CD19 (BD Biosciences), or APC‐conjugated anti‐CD45 (BD Biosciences); these are negative markers for MSCs. Propidium iodide was used to identify and exclude dead cells using flow cytometry, as previously described.16
Fitc conjugated anti human cd34
Manufactured by BD
Sourced in United States
FITC-conjugated anti-human CD34 is a fluorescently-labeled monoclonal antibody that binds to the CD34 antigen expressed on human hematopoietic stem and progenitor cells. This product is designed for use in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Fitc conjugated anti human cd44, by BD (5 mentions)
Fitc conjugated anti human cd90, by BD (3 mentions)
Pe conjugated anti hla abc, by BD (3 mentions)
Fitc conjugated anti hla dr, by BD (3 mentions)
Pe conjugated anti human cd 11b, by BD (3 mentions)
Pe conjugated anti human cd19, by BD (3 mentions)
Apc conjugated anti cd45, by BD (3 mentions)
Pe conjugated anti human cd105, by BD (3 mentions)
Fitc conjugated anti human cd29, by BD (2 mentions)
Fitc conjugated anti human cd45, by BD (2 mentions)
Apc conjugated anti human cd73, by BD (2 mentions)
Pe conjugated anti mouse cd206, by Thermo Fisher Scientific (2 mentions)
Apc conjugated anti mouse cd86, by BioLegend (2 mentions)
Human msc analysis kit, by BD (1 mentions)
Fitc conjugated anti human cd105, by BD (1 mentions)
Fitc conjugated anti human cd31, by BD (1 mentions)
Facsdiva software, by BD (1 mentions)
Lsr 2, by BD (1 mentions)
Apc conjugated anti human cd45, by BD (1 mentions)
Facscanto 2 flow cytometer, by BD (1 mentions)
7 protocols using fitc conjugated anti human cd34
1
Characterizing Mesenchymal Stem Cells by Flow Cytometry
2
Immunophenotyping of hDPSCs by Flow Cytometry
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The hDPSCs were collected in cold Dulbecco's phosphate-buffered saline (DPBS, Thermo Fisher Scientific) and subsequently incubated in the dark at 4°C for 30 min with FITC-conjugated anti-human CD29, FITC-conjugated anti-human CD44, FITC-conjugated anti-human CD105, FITC-conjugated anti-human CD31, FITC-conjugated anti-human CD34, and FITC-conjugated anti-human CD45 (BD Biosciences, San Jose, CA, USA). The samples were read on a flow cytometer (Beckman Coulter, FC500, FL, USA), and the data were analyzed using FlowJo software (Tree Star, San Carlos, CA, USA).
3
Quantification of Endothelial Progenitor Cells
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The number of EPCs was tested by fluorescence-activated cell sorting (BD LSR II with FACS Diva Software, BD Biosciences, USA). Two hundred microliters of whole blood was transferred to polystyrene tubes for testing with the following antibodies added to the samples: APC-conjugated anti-human CD45, FITC-conjugated anti-human CD34 (BD Biosciences, USA), and PE-conjugated anti-human KDR/VEGF-2 (BD Biosciences, USA). The experimental protocol can be found elsewhere (13 (link)).
4
Multiparametric Flow Cytometry Analysis
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The MSCs were stained using the following mouse monoclonal antibodies: fluorescein isothiocyanate (FITC)-conjugated anti-human CD90, phycoerythrin (PE)-conjugated anti-human CD105, allophycocyanin (APC)-conjugated anti-human CD73, FITC-conjugated anti-human CD44 (BD Biosciences), PE-conjugated anti-HLA-ABC (BD Biosciences), FITC-conjugated anti-HLA-DR (BD Biosciences), FITC-conjugated anti-human CD34 (BD Biosciences), PE-conjugated anti-human CD 11b (BD Biosciences), PE-conjugated anti-human CD 19 (BD Biosciences), and APC-conjugated anti-CD45 (BD Biosciences). Propidium iodide was used to identify and exclude the dead cells. The stained cells were acquired with a FACS Canto II flow cytometer (BD Biosystems) and analyzed using the FlowJo software (Tomy Digital Biology, Co. Ltd.). As for microglia, EYFP-expressing microglia were stained using APC-conjugated anti-mouse CD86 (BioLegend), PE-conjugated anti-mouse CD206 (ThermoFisher), and the Live/Dead™ fixable near-IR dead cell stain kit (Invitrogen). The stained cells were acquired with a Gallios flow cytometer (Beckman Coulter) and analyzed using the FlowJo software (Tomy Digital Biology, Co. Ltd.). Three independent FACS experiments were performed.
5
Flow Cytometry Immunophenotyping of Cells
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Cells were stained in FACS sorting buffer with specific antibodies for 30 min at 4 °C. The following antibodies were used: BV421-conjugated anti-human CD45 (BD Biosciences, 563879), FITC-conjugated anti-human CD34 (BD Biosciences, 555821), APC-Cy7-conjugated anti-human CD235a (BioLegend, 349116), BV605-conjugated anti-human CD44 (BD Biosciences, 562991) and PerCP-Cy5.5-conjugated 7-AAD (eBioscience, 00-6993-50). After staining, cells were washed once and resuspended in FACS sorting buffer. Cells were sorted on BD FACS Aria II. Pre-gating was first done for live cells based on a 7-AAD staining. Data analysis was performed using FlowJo V10 software (https://www.flowjo.com ).
6
Phenotypic Characterization of hDPSCs
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The hDPSCs were harvested in Dulbecco’s phosphate-buffered saline (DPBS, Thermo Scientific) and incubated for 30 min at 4 °C with FITC-conjugated anti-human CD29, FITC-conjugated anti-human CD44, FITC-conjugated anti-human CD34, and FITC-conjugated anti-human CD45 (BD Biosciences) protected from light. The samples were tested in the flow cytometer (Beckman Coulter, FC500, FL, USA), and the data were analyzed by FlowJo software (Tree Star, San Carlos, CA, USA).
7
Characterization of MSCs and Microglia
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The MSCs were stained using the following mouse monoclonal antibodies: uorescein isothiocyanate (FITC)-conjugated anti-human CD90, phycoerythrin (PE)-conjugated anti-human CD105, allophycocyanin (APC)-conjugated anti-human CD73, FITC-conjugated anti-human CD44 (BD Biosciences), PE-conjugated anti-HLA-ABC (BD Biosciences), FITC-conjugated anti-HLA-DR (BD Biosciences), FITC-conjugated antihuman CD34 (BD Biosciences), PE-conjugated anti-human CD 11b (BD Biosciences), PE-conjugated antihuman CD 19 (BD Biosciences), and APC-conjugated anti-CD45 (BD Biosciences). Propidium iodide was used to identify and exclude the dead cells. The stained cells were acquired with a FACS Canto II ow cytometer (BD Biosystems) and analyzed using the FlowJo software (Tomy Digital Biology, Co. Ltd.). As for microglia, EYFP-expressing microglia were stained using APC-conjugated anti-mouse CD86 (BioLegend), PE-conjugated anti-mouse CD206 (ThermoFisher), and the Live/Dead™ xable near-IR dead cell stain kit (Invitrogen). The stained cells were acquired with a Gallios ow cytometer (Beckman Coulter) and analyzed using the FlowJo software (Tomy Digital Biology, Co. Ltd.). Three independent FACS experiments were performed.
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