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A simple culture method for liver and intestinal tissue-resident macrophages from neonatal mice

Shimizu, Yu, Sakuragi, Naoya, Nakamura, Kiminori, Taira, Toshio, Ayabe, Tokiyoshi, Fukui, Akimasa
In vitro cellular & developmental biology 2019 v.55 no.6 pp. 436-444
coculture, cytokines, gene expression regulation, in vitro studies, inflammation, intestines, liver, macrophages, neonates, phagocytosis, phenotype, tissue repair
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.