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Cercarial Transformation and in vitro Cultivation of Schistosoma mansoni Schistosomules
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Schistosomiasis and in vitro transdifferentiation of murine peritoneal macrophages into fibroblastic cells
M Godoy1, M Geuskens, E A Van Marck
1Department of Molecular Biology, Free University of Brussels, Rhode-St-Genèse, Belgium.
Abstract:
We developed a method for avoiding contamination by fibroblasts when cultures of peritoneal cells are initiated. Macrophages were identified by immunogold detection [light microscope, transmission (TEM) and scanning (SEM) electron microscopes] of membrane antigens (Mac-1+, Thy-1,2-), non-specific esterase activity and ultrastructural features (TEM). As compared with controls, the yield of peritoneal macrophages was 2- and 12-fold higher, respectively, in acutely and chronically infected mice. In all, 30 "chronic", 18 "acute" and 18 control cultures were followed up. At a given cell-density seeding, the decline of control, "acute" and "chronic" cultures starts at about day 10, 15, and 27, respectively. In "chronic" cultures only, fibroblast-like cells appear from day 6 onwards; their number increases with time. Cells showing characters intermediary between macrophages and fibroblasts were observed. We suggest that fibroblast-like cells result from the in vitro transdifferentiation of a limited number of in vivo committed macrophages.
Insights
Researchers found that fibroblast-like cells in peritoneal cell cultures may originate from macrophages. This suggests a potential in vitro transdifferentiation process, impacting macrophage culture yields and longevity.
Area of Science:
- Immunology
- Cell Biology
- Microscopy
Background:
- Contamination by fibroblasts can complicate the initiation and study of peritoneal cell cultures.
- Macrophages are key immune cells within peritoneal exudates, crucial for various biological processes.
- Understanding macrophage behavior in vitro is essential for immunological research.
Purpose of the Study:
- To develop a method for preventing fibroblast contamination in peritoneal cell cultures.
- To investigate the origin and behavior of fibroblast-like cells appearing in these cultures.
- To analyze the yield and longevity of peritoneal macrophages under different experimental conditions.
Main Methods:
- Macrophages were identified using immunogold detection of membrane antigens (Mac-1+, Thy-1,2-), non-specific esterase activity, and transmission electron microscopy (TEM) ultrastructural features.
- Peritoneal cells were cultured from acutely and chronically infected mice, compared to controls.
- Cultures were monitored for cell viability, fibroblast appearance, and intermediary cell types over time.
Main Results:
- Macrophage yields were significantly higher (2-fold in acute, 12-fold in chronic infection) compared to controls.
- Fibroblast-like cells emerged in 'chronic' cultures from day 6 onwards, increasing with time.
- Intermediary cells with macrophage and fibroblast characteristics were observed, suggesting transdifferentiation.
Conclusions:
- Fibroblast-like cells in peritoneal cultures likely arise from the in vitro transdifferentiation of committed macrophages.
- This transdifferentiation process may influence the observed differences in culture longevity between control, acute, and chronic groups.
- The findings offer insights into macrophage plasticity and potential strategies for optimizing peritoneal cell cultures.
