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Published on: November 3, 2014
Transcriptomic Analysis of Rat Macrophages
Clare Pridans1,2, Katharine M Irvine3, Gemma M Davis4
1Centre for Inflammation Research, University of Edinburgh Centre for Inflammation Research, Edinburgh, United Kingdom.
Abstract:
The laboratory rat is widely used as a model for human diseases. Many of these diseases involve monocytes and tissue macrophages in different states of activation. Whilst methods for in vitro differentiation of mouse macrophages from embryonic stem cells (ESC) and bone marrow (BM) are well established, these are lacking for the rat. The gene expression profiles of rat macrophages have also not been characterised to the same extent as mouse. We have established the methodology for production of rat ESC-derived macrophages and compared their gene expression profiles to macrophages obtained from the lung and peritoneal cavity and those differentiated from BM and blood monocytes. We determined the gene signature of Kupffer cells in the liver using rats deficient in macrophage colony stimulating factor receptor (CSF1R). We also examined the response of BM-derived macrophages to lipopolysaccharide (LPS). The results indicate that many, but not all, tissue-specific adaptations observed in mice are conserved in the rat. Importantly, we show that unlike mice, rat macrophages express the CSF1R ligand, colony stimulating factor 1 (CSF1).
Insights
Researchers developed methods to create rat macrophages from embryonic stem cells (ESC) and bone marrow (BM). Rat macrophages share similarities with mouse macrophages but have unique gene expression profiles, including expressing colony stimulating factor 1 (CSF1).
Area of Science:
- Immunology
- Stem Cell Biology
- Comparative Genomics
Background:
- Laboratory rats are crucial models for human diseases involving monocytes and macrophages.
- Established methods for mouse macrophage differentiation from ESC and BM exist, but are lacking for rats.
- Rat macrophage gene expression profiles are less characterized than mouse counterparts.
Purpose of the Study:
- To establish methodology for producing rat ESC-derived macrophages.
- To compare gene expression profiles of rat macrophages from various sources (ESC, BM, lung, peritoneal, blood).
- To characterize Kupffer cells and rat BM-derived macrophage responses to LPS.
Main Methods:
- Differentiation of rat macrophages from embryonic stem cells (ESC) and bone marrow (BM).
- Gene expression profiling of various rat macrophage populations.
- Analysis of Kupffer cells in CSF1R-deficient rats.
- Lipopolysaccharide (LPS) stimulation of BM-derived macrophages.
Main Results:
- Successful establishment of rat ESC-derived macrophage production methodology.
- Comparison of gene expression profiles revealed conserved and divergent tissue-specific adaptations compared to mice.
- Rat macrophages express CSF1, the ligand for CSF1R, which is not typically observed in mice.
Conclusions:
- The study provides a foundational methodology for generating rat macrophages in vitro.
- Rat macrophages exhibit conserved and unique characteristics compared to mouse macrophages, impacting their utility as disease models.
- The expression of CSF1 in rat macrophages suggests distinct regulatory mechanisms compared to mice.
