NP cells at a density of 1 × 105 cells per well were seeded into six-well plates. An Annexin V/PI apoptosis detection kit (BD Biosciences, Franklin Lakes, NJ, USA) was used to determine the apoptotic rate of cells. NP cells were washed twice with cold PBS, suspended in 400 ul Annexin V binding buffer (1X) and stained with 5 μl Annexin V-FITC and 10 μl PI for 15 min in the dark. The stained cells were analyzed using the FACScan flow cytometry system (Beckman Coulter, CA, USA)
Facscan flow cytometry system
Manufactured by Beckman Coulter
Sourced in United States
The FACScan flow cytometry system is a compact and versatile instrument designed for multi-parameter analysis and sorting of individual cells or particles. It utilizes laser-based detection technology to precisely measure and characterize various physical and chemical properties of cells, including size, granularity, and fluorescence intensity.
Automatically generated - may contain errors
2 protocols using facscan flow cytometry system
1
Apoptosis Evaluation of NP Cells
2
Isolation and Characterization of Rabbit Bone Marrow Mesenchymal Stem Cells
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A density gradient centrifugation approach was used to collect mononuclear cells from the rabbit bone marrow with the Percoll cell separation solution (Solarbio, China), after which these cells were cultured in T25 cell culture flasks containing αMEM supplemented with 10% fetal bovine serum (FBS; Gibco, USA) and 1% penicillin-streptomycin (Gibco, USA) at 37°C in a humidified 5% CO2 incubator. Media were changed every 3 days, and rBMSCs from passages 3–5 were used for all experiments. The surface phenotype of these rBMSCs was assessed via flow cytometry by staining 5 × 105 BMSCs from passage 3 with antibodies specific for CD34 (1 : 1000; BioLegend, CA, USA), CD45 (1 : 1000; BioLegend), CD90 (1 : 500; BioLegend), CD105 (1 : 1000; BioLegend), HLA-DR (1 : 1,000; Abcam, Cambridge, UK), and CD44 (1 : 1,000; Abcam) for 30 minutes at 4°C while protected from light. Cells were then rinsed with PBS and analyzed using a FACScan flow cytometry system (CytoFLEX; Beckman Coulter, USA).
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