Related Experiment Video
Updated: Aug 8, 2026
Use of a Monocyte Monolayer Assay to Evaluate Fcγ Receptor-mediated Phagocytosis
Published on: January 2, 2017
[Attempt to use monocytes for HLA-DR typing]
Abstract:
Two methods of isolating monocytes from peripheral defibrinized blood are described and compared with each other: 1. Adherence of monocytes on the surface of Petri-dishes and 2. Two-phase technique with centrifugation over a gradient of Percoll. The technique with Percoll has advantages as compared with the method of adherence: 1. Greater purity of monocyte suspension and higher percentage of monocytes (85% of monocytes on an average compared with 60% in adherence). 2. A damage of membrane can sometimes be observed in those monocytes gained by adherence and as a consequence of that there is a tendency to unspecifically positive cytotoxic reactions. In monocytes isolated by means of these two methods HLA-DR antigens could be identified. There was no complete correspondence between the cytotoxical reactions with monocytes and B-lymphocytes.
Insights
The Percoll gradient technique yields higher purity and percentage of monocytes compared to the adherence method. This improved monocyte isolation method is crucial for accurate immunological studies.
Area of Science:
- Immunology
- Cell Biology
Context:
- Peripheral blood mononuclear cells (PBMCs) are vital for immunological research.
- Accurate isolation of specific cell types, like monocytes, is essential for reliable experimental outcomes.
Purpose:
- To compare the efficacy of two monocyte isolation methods: adherence to Petri dishes and a two-phase Percoll gradient centrifugation.
- To evaluate the purity, monocyte yield, and cellular integrity of monocytes obtained by each method.
Summary:
- The Percoll gradient technique provides a higher monocyte purity (average 85%) and yield compared to the adherence method (average 60%).
- Monocytes isolated via adherence may exhibit membrane damage, leading to non-specific cytotoxic reactions, unlike those from the Percoll method.
- Both isolation methods allowed for the identification of Human Leukocyte Antigen - antigen D Related (HLA-DR) on monocytes.
Impact:
- The Percoll method offers a superior approach for obtaining high-quality monocytes for immunological assays.
- This research aids in selecting optimal cell isolation protocols, enhancing the reproducibility and validity of immunological studies.
- Findings contribute to a better understanding of monocyte characteristics and their role in immune responses.
