A human macrophage cell line (human CD14+ monocytes from peripheral blood, single donor; C-12909, http://www.promocell.com/product/human-cd14-monocytes-hmocd14-pb/ ) from PromoCell GmbH (Heidelberg, Germany) was cultured at a density of 1×106 cells in Monocyte Attachment Medium (PromoCell GmbH). Cells were cultured in M1-Macrophage Generation Medium DXF so that they could differentiate into M1- and M2-like macrophages as in our previous study (19 (link)). In addition, these cells were treated with 1, 10 or 20 ng/ml of interleukin-4 (IL-4; R&D Systems) for 24 h. Human monocytic (THP-1) cells were obtained from the American Type Culture Collection and maintained in RPMI-1640 containing 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.), 1% penicillin-streptomycin (Gibco; Thermo Fisher Scientific, Inc.), and 2 mmol/l of glutamine (Gibco; Thermo Fisher Scientific, Inc.).
Protocol detail
Macrophage Polarization with IL-4
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C-12909
Monocyte Attachment Medium
IL-4
Human monocytic (THP-1) cells
RPMI-1640
fetal bovine serum
penicillin-streptomycin
glutamine
Cell line
Cells
Cultured cells
Cultured medium
Donor
Fetal bovine serum
Glutamine
Human
Interleukin 20
Macrophages
Monocytic
Penicillin
Streptomycin
Thp 1 cells
Corresponding Organization :
Other organizations : Kitakyushu City Yahata Hospital, University of Occupational and Environmental Health Japan
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Variable analysis
independent variables
- Interleukin-4 (IL-4) concentration (1 ng/ml, 10 ng/ml, 20 ng/ml)
- Differentiation of monocytes into M1- and M2-like macrophages
- Human CD14+ monocytes from peripheral blood, single donor (C-12909, PromoCell)
- Monocyte Attachment Medium (PromoCell GmbH)
- M1-Macrophage Generation Medium DXF
- Human monocytic (THP-1) cells (American Type Culture Collection)
- RPMI-1640 medium containing 10% fetal bovine serum, 1% penicillin-streptomycin, and 2 mmol/l of glutamine
- Positive control: Differentiation of monocytes into M1- and M2-like macrophages without IL-4 treatment, as in the previous study (19)
- Negative control: Not explicitly mentioned
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