The Effect of Aspirin and Ibuprofen on the Phagocytic Ability of Macrophages

Abstract

Source:
Adepemi O. Ogundeji¹, Nozethu Mjokane¹, Maphori Maliehe¹, Olihile M. Sebolai<sup>1

1</sup>Department of Microbiology and Biochemistry, University of the Free State.

This video showcases a method to assess the impact of aspirin and ibuprofen, at their minimum inhibitory concentrations, on macrophage phagocytosis using pH-sensitive fluorescent dye-stained fungal cells. The drugs inhibit cyclooxygenase-2, decreasing prostaglandin formation and boosting macrophage phagocytosis, as observed through fluorescence measurements of pH-sensitive dye.

Protocol

1. Safety considerations

1. Personnel should use personal protective equipment (PPE).
2. Handle the pathogen and macrophages in a Class II Biological safety cabinet.

2. Preparation of macrophages

1. Cultivate a murine macrophage cell line, RAW 264.7, in a tissue flask containing 10 mL of Roswell Park Memorial Institute-1640 (RPMI-1640) medium, supplemented with 20 mg/mL streptomycin, 2 mM L-glutamine, 20 U/mL penicillin and 10% fetal bovine serum (FBS).
2. Place the flask in a 5% carbon dioxide (CO₂) incubator set at 37˚C until 80% confluence is achieve....