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Evaluation of various filter membranes for Boyden-type leukocyte migration studies
Abstract:
Various types and makes of membranes were assessed for suitability in Boyden-type chemotaxis studies. Pooled exudate neutrophils and macrophages from the rat were used as indicator cells. Sartorius filters were found to be superior to other cellulose ester filters both in efficiency and consistency of cell migration. Variations between and within Sartorius filter lots were small and filter inconsistency could be ruled out as a factor in day-to day variation in efficiency of cell migration. In our hands, Millipore membranes were unsatisfactory for chemotaxis studies. With the exception of the shortened incubation times, polycarbonate filters did not offer significant advantages over Sartorius filters when the vigorously responding rat exudate cells were tested. The orientation of the cellulose ester filters in the chemotaxis chambers was found to be a critical factor when the lowermost level counting method was used. The observed polarity could not readily be washed out by acid, alkali or ethanol treatment nor by boiling the filters in distilled water and remains unexplained.
Insights
Sartorius filters are superior for Boyden-type chemotaxis studies, showing consistent cell migration. Millipore membranes were unsatisfactory, while polycarbonate filters offered no significant advantages over Sartorius filters for rat exudate cells.
Area of Science:
- Immunology
- Cell Biology
Background:
- Boyden-type chemotaxis assays are crucial for studying cell migration.
- Membrane selection impacts the reliability and reproducibility of chemotaxis results.
Purpose of the Study:
- To evaluate the suitability of various membranes for Boyden-type chemotaxis.
- To identify optimal membrane materials for consistent neutrophil and macrophage migration.
Main Methods:
- Assessed multiple membrane types (Sartorius, Millipore, polycarbonate) using rat exudate neutrophils and macrophages.
- Conducted Boyden-type chemotaxis assays to measure cell migration efficiency and consistency.
- Investigated the impact of filter orientation on cell migration.
Main Results:
- Sartorius cellulose ester filters demonstrated superior efficiency and consistency compared to other cellulose ester filters.
- Millipore membranes proved unsatisfactory for chemotaxis studies.
- Polycarbonate filters showed no significant advantage over Sartorius filters for vigorous rat exudate cells.
- Filter orientation was critical for cell migration when using the lowermost level counting method.
Conclusions:
- Sartorius filters are recommended for reliable Boyden-type chemotaxis studies.
- Filter consistency and orientation are critical factors influencing cell migration assays.
- Further investigation is needed to explain the observed unexplained filter polarity.