Use of a microtitre plate chemiluminescence reader to study surface phagocytosis by human monocytes
✍ Scribed by Cree, Ian A. ;Blair, Andrea L. ;Beck, J. Swanson
- Publisher
- John Wiley and Sons
- Year
- 1989
- Weight
- 252 KB
- Volume
- 3
- Category
- Article
- ISSN
- 0884-3996
No coin nor oath required. For personal study only.
✦ Synopsis
DD1 9SY. UK Human mononuclear cells were separated from freshly obtained peripheral venous blood by density centrifugation and the number of monocytes present estimated by volume spectroscopy. The mononuclear cells were then placed directly into the wells of a microtitre plate and incubated for one hour at 37°C t o promote adherence of the monocytes t o the plastic wells. Non-adherent cells were then removed by washing, thus avoiding the need t o treat the monocytes with EDTA or other reagents during cell preparation. The time course and dynamics of the chemiluminescence response of adherent monocytes towards opsonized zymosan was similar t o those seen using non-adherent cells.
The ability of adherent monocyte preparations t o produce chemiluminescence following incubation for varying periods with T-lymphocyte conditioned medium was investigated. The use of a microtitre plate chemiluminescence reader allows several plates t o be assayed over the 24-hour period and since small numbers of cells are required, many cultures can be analysed in one experiment. This technique (Patent applied for) promises t o be a powerful tool for dissecting the cellular events which occur during macrophage activation and examining the effect of various lymphokines on the ability of monocytes t o produce a chemiluminescence response.