THP-1 monocytic cells (Sigma-Aldrich, 100 μl of 5 × 106/ml) were seeded onto 96 well plates in 10% FBS (EuroClone) supplemented Dulbecco’s Modified Eagle Medium (DMEM, Lonza) and incubated overnight at 37 °C, with 5% CO2. Infection of monocytes with C. parapsilosis CDC 317 cells was performed in 1:5 ratio. Cells were co-incubated at 37 °C with 5% CO2 and collected by centrifugation (1000 rpm, 5 min) at 1 or 6 h post-infection. Cells were washed once with ice-cold PBS and pellet was suspended in RNase-free distilled water. For fungal RNA isolation, monocytes were lysed through 27 G needles and RNA extraction was performed with the RNeasy Plant Mini Plus Kit (Qiagen) according to manufacturer’s instructions.
Thp 1 monocytic cells
Manufactured by Merck Group
Sourced in Germany, United States
THP-1 monocytic cells are a commercially available human cell line derived from an acute monocytic leukemia patient. They are commonly used in cell biology research as a model for studying monocyte and macrophage functions.
Automatically generated - may contain errors
Lab products found in correlation
Rneasy plant mini plus kit, by Qiagen (1 mentions)
Fetal bovine serum (fbs), by Euroclone (1 mentions)
Dulbecco modified eagle medium (dmem), by Lonza (1 mentions)
Protein g sepharose 4 fast flow, by Cytiva (1 mentions)
Hek293 cells, by Merck Group (1 mentions)
Expi293f, by Thermo Fisher Scientific (1 mentions)
Thp 1 monocytic cells, by American Type Culture Collection (1 mentions)
Phorbol 12 myristate 13 acetate, by Merck Group (1 mentions)
Hek293 cell, by American Type Culture Collection (1 mentions)
4 protocols using thp 1 monocytic cells
1
Infecting THP-1 Monocytes with C. parapsilosis
2
Antibody Production in Cell Lines
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THP-1 monocytic cells (Sigma Aldrich, 88081201-1VL) were cultured in RPMI with 10% FBS and L-glutamine and were kept at 2.5-10 × 105 cells per mL. HEK293 cells (Sigma Aldrich, 12022001‐1VL) and Expi293F (Thermofisher, A14527) cells were used to produce antibodies similarly as done before37 (link),38 (link). Supernatant containing produced antibodies was purified through incubation with protein G sepharose 4 fast flow (Cytiva, 17-0618-05) according to the manufacturer’s instruction.
3
Differentiation of THP-1 Monocytes to Macrophages
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To establish an inflammatory model, THP-1 monocytic cells (ATCC, USA) were seeded into cell culture flasks. By adding 100 nM phorbol-12-myristate 13-acetate (Sigma-Aldrich, Germany), THP-1 monocytic cells were differentiated into macrophages for 3 days. Then, the PMA-containing medium was removed. The differentiation of PMA-treated THP-1 cells was extended by incubating the cells in fresh culture medium for an additional 5 days. Cells that continued to receive regular growth medium were designated as nonactivated macrophages (NM).
4
Differentiation of THP-1 Macrophages into Foam Cells
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HEK293 cells were obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA) and cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS). The hum an THP-1 monocytic cell line (ATCC) was cultured in complete RPMI 1640 medium that was supplemented with 10% heat-inactivated FBS. Phorbol 12-myristate 13-acetate (PMA, 160 nM) (Sigma, St. Louis, MO, USA) was added to the media, and THP-1 monocytic cells were incubated for 72 h to induce a macrophage phenotype of differentiation. These cells were incubated in a humidified incubator at 37 ℃ with 5% CO2. To fully differentiate macrophages into foam cells and clearly observe the changes of UFM1 expression in foam cell formation, macrophages were first cultured in RPMI 1640 medium (serum free) containing different concentrations of oxLDL for 24 h or exposed to 50 μg/ mL of oxLDL for indicated times. In subsequent experiments, THP-1 stable transgenic macrophages (LV-UFM1 and sh-UFM1) were cultured in complete RPMI 1640 medium containing 50 μg/mL oxLDL for indicated times.
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