RRC ID |
57802
|
著者 |
Shimizu Y, Sakuragi N, Nakamura K, Taira T, Ayabe T, Fukui A.
|
タイトル |
A simple culture method for liver and intestinal tissue-resident macrophages from neonatal mice.
|
ジャーナル |
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.
|
巻・号 |
55(6)
|
ページ |
436-444
|
公開日 |
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 |
1.665
|
引用数 |
0
|
リソース情報 |
ヒト・動物細胞 |
NIH3T3-3-4(RCB1862) |