Protocol for isolating and culturing human monocytes from peripheral blood and measuring intracellular calcium dynamics using Fura-2

Key Points

• This research aims to outline a reliable protocol for isolating human monocytes and analyzing their intracellular calcium levels.
• Isolated human monocytes from peripheral blood through density gradient centrifugation.
• Cultured monocytes in appropriate media under sterile conditions.
• Measured intracellular calcium levels using Fura-2 dye and fluorescence microscopy.
• Successful isolation of viable human monocytes with high purity rates.
• Demonstrated the effective measurement of intracellular calcium dynamics using Fura-2.
• Established a feasible protocol for future studies on calcium signaling in immune cells.