Confluent HUVECs cultured in 24‐well plates were incubated at 37°C in a 5% CO2 gassed incubator for 16 hours with EGM‐2 containing 0.5% FBS for 16 hours and then treated with the indicated concentrations of KP‐10 for 4 hours. Subsequently, human primary monocytes or THP‐1 cells (Health Science Research Resources Bank, Osaka, Japan) were labeled with Cell trace calcein red‐orange (Life Technologies, Carlsbad, CA) with 1×105 cells added to each well of a HUVEC‐seeded 24‐well plate. After 1 hour, cells were washed 4 times, with human primary monocytes or THP‐1 cells bound to HUVECs, and then examined by fluorescence microscopy (IX70; Olympus, Tokyo, Japan). Their adhesion was assessed using image analysis software (ImageJ; National Institutes of Health, Bethesda, MD).
Thp 1 cells
Manufactured by Olympus
Sourced in Japan
THP-1 cells are a human monocytic cell line derived from an acute monocytic leukemia patient. They are widely used in immunological and cell biology research as a model for studying monocyte and macrophage function.
Automatically generated - may contain errors
Lab products found in correlation
Calcein am solution, by Merck Group (2 mentions)
Thp 1 cells, by RIKEN BioResource Center (2 mentions)
Celltrace calcein red orange, by Thermo Fisher Scientific (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
Thp 1 cells, by Merck Group (1 mentions)
Fluorescence microscopy, by Olympus (1 mentions)
3 protocols using thp 1 cells
1
Quantifying HUVEC-Monocyte Adhesion
2
Casticin Attenuates TNF-α Induced Monocyte-Epithelial Adhesion
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Human bronchial epithelial BEAS-2B cells were cultured in DMEM/F12 and seeded into 6-well plates. BEAS-2B cells were pre-treated with casticin (5–20 μM) for 1 h, and then incubated with 10 ng/ml TNF-α for 24 h as described previously (Huang et al., 2016 (link)). Human monocytic THP-1 cells (5 × 106/ml) (Bioresource Collection and Research Center, Taiwan) were incubated with calcein-AM solution (Sigma) for 0.5 h. Next, THP-1 cells were co-cultured with BEAS-2B cells and observed under a fluorescence microscope (Olympus).
3
Immune Cell Adhesion Assay for BEAS-2B Cells
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BEAS-2B cells were treated with various doses of HZW or HZE for 1 h and then stimulated with TNF-α/IL-4 for 24 h in 6-well plates as described previously [30 (link), 31 (link)]. THP-1 cells (Bioresource Collection and Research Center, Taiwan) were treated with calcein-AM solution (Catalog No. C1359; Sigma) at 37°C for 30 min, and THP-1 cells were cocultured with BEAS-2B cells at 37°C for 30 min. Finally, THP-1 cells adhered to BEAS-2B cells were measured using fluorescence microscopy (Olympus, Tokyo, Japan).
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