A simple culture method for liver and intestinal tissue-resident macrophages from neonatal mice
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The liver and intestine contain a remarkably large portion of tissue-resident macrophage cells representing a phenotype that downregulates inflammation and initiates tissue repair. Here, liver and intestinal tissues obtained from neonatal mice were minced, enzymatically digested, and incubated in RPMI1640-based media. In a 2-wk culture, spherical floating cells emerged on a fibroblastic sheet. These cells showed phagocytic activity and F4/80+-CD11b+-CD206+-Arg1+-iNOS−-CD209a− phenotype, suggesting that these cells are tissue-resident macrophages. These macrophages proliferated in the co-culture system in the presence of fibroblastic feeder cell layer and absence of supplemental cytokines; the co-culture system did not cause a significant change in the phenotype of cells grown in a 4-wk culture. On the feeder cells, macrophage density was approximately 1.5 × 104/cm2 and the doubling time was approximately 70 h. Based on these observations, we present a simple method for the isolation and propagation of tissue-resident macrophages resembling M2 macrophage from neonatal mice, and this method provides a useful platform for in vitro studies of tissue-resident macrophages.
KeywordsIntestinal macrophage Tissue-resident macrophage Simple culture method Feeder cell
The authors thank Kazuya Furusawa and Naoki Sasaki for the discussions and suggestions.
This work was supported by Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (No. 17K11661 to KN and 18H02788 to TA), the Center of Innovation Program from the Japan Science and Technology Agency (to NS, KN, and TA), and a Chuo University Personal Research Grant (to AF).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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