Approximately 1 × 106 cervical cancer cells per condition were seeded onto Petri dishes. Cells were incubated for 35 min at 4°C in staining buffer (PBS, 0.5% BSA, and 0.01% sodium azide). The cells were stained with specific antibodies against the following surface markers: anti-NKp30-PE, anti-CD95-APC, anti-CD39-FITC, anti-CD-73-PE-Cy7, anti-KIR3DL1-APC, anti-NKp46-APC, anti-CD25-PE, anti-CD25-PErCP-Cy5.5, anti-CD44-FITC, and anti-CD24-PE (Becton Dickinson, USA); anti-NKG2A-PerCP, anti-CD158e1-APC (R&D, Minneapolis, MN); and anti-CD158b2-FITC (BioLegend). Cells were washed and fixed with 1% paraformaldehyde for 20 minutes. For permeabilisation, cells were treated with Cytofix/Cytoperm buffer for 20 minutes (Becton Dickinson, USA). For intracellular staining, anti-human CTLA-4-APC, anti-CD95-PE-Cy7 (Becton Dickinson, USA), and anti-NKG2A-PerCP (R&D, Minneapolis, MN) antibodies were used. Cell samples were incubated for 35 minutes at 4°C in the dark. Finally, cells were washed and resuspended in staining buffer. The phenotype of tumor cells was characterized by flow cytometry on a FACSAria II cytometer (Becton Dickinson, USA). Data were analysed using Summit 4.4 software.
Anti cd 73 pe cy7
Manufactured by BD
Sourced in United States
Anti-CD-73-PE-Cy7 is a fluorescently-labeled antibody that binds to the cell surface protein CD73. CD73 is an enzyme involved in the regulation of extracellular adenosine levels. The PE-Cy7 fluorescent label allows for the detection and analysis of CD73-expressing cells using flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd90 apc, by BD (3 mentions)
Anti cd34 pe, by BD (2 mentions)
Facsaria 2 cytometer, by BD (1 mentions)
Anti cd24 pe, by BD (1 mentions)
Anti cd25 pe, by BD (1 mentions)
Cytofix cytoperm buffer, by BD (1 mentions)
Anti nkg2a percp, by R&D Systems (1 mentions)
Anti cd14 fitc, by BD (1 mentions)
Stempro differentiation kit, by Thermo Fisher Scientific (1 mentions)
Anti cd45 fitc, by BD (1 mentions)
Facscanto 2 flow cytometer, by BD (1 mentions)
Anti cd45 percp, by BD (1 mentions)
Cd105 pe, by BD (1 mentions)
Anti cd14 apc, by BD (1 mentions)
Hla dr pe, by BD (1 mentions)
Anti cd105 pe, by BD (1 mentions)
Flow cytometry, by BD (1 mentions)
E8 media, by Thermo Fisher Scientific (1 mentions)
Novoexpress, by Agilent Technologies (1 mentions)
Cd79a pe, by BD (1 mentions)
4 protocols using anti cd 73 pe cy7
1
Phenotyping Cervical Cancer Cells
2
MSC Immunophenotyping and Differentiation
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The Cell Therapy and Engineering Unit of EFS Auvergne-Rhône-Alpes provided pre-immunophenotyped MSCs with anti-CD45-FITC, anti-CD14-FITC, anti-CD105-PE, anti-CD90-APC and anti-CD73-PE-CY7 (Becton Dickinson Franklin Lakes, NJ, USA) which were analysed by flow cytometry according to the manufacturer’s instructions (data not shown). To confirm MSC functionality, cells were differentiated using induction media (Stempro differentiation kit, Thermo Fisher Scientific, Waltham, MA, USA) for adipocyte, chondrocyte and osteocyte formation and stained respectively with Oil Red O, Alcian blue and Alizarin Red S.
3
Flow Cytometric Analysis of ADMSC Markers
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The expression of ADMSC surface markers was assessed by flow cytometric analysis. Cells were trypsinised into a single cell suspension and incubated with the fluorochrome-conjugated antibodies against human antigens, including anti-CD34-PE, anti-CD45-PerCP, anti-CD73-PE/Cy7, anti-CD90-APC, and CD105-PE (BD Biosciences, San Jose, California), at 4°C for 30 minutes. The cells were then washed twice with cold PBS and fixed with 1% paraformaldehyde before analysis by FACSCanto II Flow Cytometer (BD Biosciences) using FACSDiva software.
4
Derivation and Characterization of ESC-Derived MSCs
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ESC-MSCs were differentiated from ESC using a two-step process. In brief, H9-ESC colonies (YuanSheng Biotech Corporation, Hangzhou, China) were dissociated into small clumps after 3 min of incubation with TrypLE Express and then transferred to ultralow-attachment plates in E8 media (Gibco, Grand Island, NY, USA). After 7 days, embryoid bodies (EBs) were harvested and plated in MSC induction medium composed of Dulbecco’s modified Eagle’s medium (high glucose), 10% fetal bovine serum, and 1 mM l -glutamine. After 2 w, the EB outgrowths were subcultured using TrypLE Express. These cells were ESC-MSCs and were designated passage 0 (P0). The differentiated ESC-MSCs attained a homogenous population of spindle-shaped cells. Passage 4 (P4) ESC-MSCs were used in the following animal experiment.
In this study, phenotypic identification of MSCs was performed using flow cytometry (BD Biosciences, USA) with the following monoclonal antibodies: anti-CD73-PE-Cy7, anti-CD90-APC, anti-CD105-PE, anti-CD14-APC, anti-CD34-PE, anti-CD45-fluorescein isothiocyanate (FITC), CD79a-PE, and HLA-DR-PE. All antibodies were purchased from BD Pharmingen (China). After antibody labeling, data were acquired using an Agilent NovoCyte and analyzed using NovoExpress.
In this study, phenotypic identification of MSCs was performed using flow cytometry (BD Biosciences, USA) with the following monoclonal antibodies: anti-CD73-PE-Cy7, anti-CD90-APC, anti-CD105-PE, anti-CD14-APC, anti-CD34-PE, anti-CD45-fluorescein isothiocyanate (FITC), CD79a-PE, and HLA-DR-PE. All antibodies were purchased from BD Pharmingen (China). After antibody labeling, data were acquired using an Agilent NovoCyte and analyzed using NovoExpress.
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