RRC ID 57802
Author Shimizu Y, Sakuragi N, Nakamura K, Taira T, Ayabe T, Fukui A.
Title A simple culture method for liver and intestinal tissue-resident macrophages from neonatal mice.
Journal In Vitro Cell Dev Biol Anim
Abstract 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.
Volume 55(6)
Pages 436-444
Published 2019-6-1
DOI 10.1007/s11626-019-00359-y
PII 10.1007/s11626-019-00359-y
PMID 31119642
MeSH Animals Animals, Newborn Cell Culture Techniques / methods* Cell Separation / methods* Coculture Techniques Female Fibroblasts / cytology Flow Cytometry Fluorescent Antibody Technique Gene Expression Regulation Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism Intestine, Small / cytology* Liver / cytology* Macrophages / cytology* Macrophages / physiology Mice, Inbred C57BL Mice, Inbred ICR Mice, Transgenic Phagocytosis
IF 1.645
Times Cited 0
Resource
Human and Animal Cells NIH3T3-3-4(RCB1862)