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A Simple and Efficient Method to Isolate Macrophages from Mixed Primary Cultures of Adult Liver Cells
Published on: May 24, 2011
Cultures of pig macrophages
Abstract:
Freshly collected blood from pigs was defibrinated with glass beads. The white cells were collected on a Ficoll-Hypaque mixture and grown in RPMI 1640 medium containing 20 per cent fetal calf serum. Monocytes adhered to the plastic or glass flasks and the erythrocytes and lymphocytes remained in suspension. The adherent cells assumed the appearance of macrophages and the mean cell size at eight days was 40 mum. Phagocytosis was demonstrated and, in the first few days, incorporation of tritiated thymidine occurred. Cells remained viable in culture for more than eight weeks. Histo-chemical studies and the appearance of the cells under the electron microscope are described.
Insights
This study details a method for isolating and culturing pig monocytes, which transform into macrophages. These macrophages exhibit phagocytosis and remain viable in culture for extended periods, offering a valuable model for immunological research.
Area of Science:
- Veterinary Immunology
- Cell Biology
- Animal Models
Background:
- Monocytes are crucial immune cells with diverse functions.
- Establishing reliable in vitro models for studying monocytes/macrophages is essential for research.
Purpose of the Study:
- To describe a method for isolating and culturing porcine monocytes.
- To characterize the resulting macrophage-like cells in vitro.
Main Methods:
- Defibrination of pig blood followed by Ficoll-Hypaque density gradient centrifugation.
- Culture of isolated white blood cells in RPMI 1640 with fetal calf serum.
- Adherence selection to isolate monocytes, with subsequent morphological and functional assessments.
Main Results:
- Isolated monocytes adhered to culture flasks and differentiated into macrophage-like cells within days.
- Macrophage-like cells reached a mean size of 40 µm by day eight and demonstrated phagocytic activity.
- Cells maintained viability in culture for over eight weeks, with described histo-chemical and ultrastructural features.
Conclusions:
- A robust method for generating pig macrophage cultures from peripheral blood monocytes was established.
- These cultured macrophages provide a viable and functional model for studying porcine immunology and cellular processes.
