THP-1 cells stably transduced with GFP-actin were maintained in a suspension culture in RPMI media (Gibco, Grand Island, NY, USA), supplemented with 10% FBS (HyClone, Rockford, IL, USA), 1% penicillin streptomycin (Invitrogen, Carlsbad, CA, USA), and 0.05 mM 2-mercaptoethanol. [30 ,36 (link)] For all of the experiments, the cells (13,000 cells/cm2) were plated onto glass coverslips (Ted Pella Inc.) in 35 mm polystyrene petri dishes (BD, Ashland, MA, USA). The THP-1 cells were stimulated with the addition of 1 μg/mL of Phorbol 12-myristate 13-acetate, PMA, for 72 h.
35 mm polystyrene petri dishes
Manufactured by BD
Sourced in United States
35 mm polystyrene petri dishes are a type of sterile, disposable laboratory equipment used for the culture and observation of microorganisms. They provide a controlled environment for the growth and study of various cell types.
Automatically generated - may contain errors
Lab products found in correlation
Glass coverslips, by Ted Pella (4 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Rpmi media, by Thermo Fisher Scientific (2 mentions)
Mycoalert kit, by Lonza (2 mentions)
Vascu life vegf cell culture media, by Lifeline Cell Technology (2 mentions)
Adult human dermal fibroblasts, by Lifeline Cell Technology (2 mentions)
Fibrolife cell culture media, by Lifeline Cell Technology (2 mentions)
4 protocols using 35 mm polystyrene petri dishes
1
Differentiation of GFP-actin THP-1 Cells
2
Endothelial and Fibroblast Cell Culture
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Human vascular endothelial cells were cultured in Vascu-Life VEGF Cell Culture Media (Lifeline Cell Technology, Walkersville, MD) and checked for mycoplasma contamination using MycoAlert Kit (Lonza, Rockland, ME). Adult human dermal fibroblasts (Lifeline Cell Technology, Walkersville, MD) were cultured in Fibrolife cell culture media (Lifeline Cell Technology, Walkersville, MD), as previously dsescribed. [39 ] The cells were plated onto glass coverslips (Ted Pella Inc.), coated with fibronectin and plated at a density of 5000 cells/cm2 in 35 mm polystyrene petri dishes (BD, Ashland, MA, USA). Prior to the experiments, the cells were allowed to grow for 48 h. The AFM analysis was carried out at room temperature (22°C) in HBSS, pH 7.4 with 1.3 mmol/l CaCl2, 0.9 mmol/l MgCl2, 2 mmol/l GlutaMax, 10 g/l heparin, 5.6 mmol/l glucose, and 1% FBS) analogous to C. M. Coll-Ferrer et al. [30 ]
3
Endothelial Cell Culture and AFM Analysis
4
Differentiation of GFP-actin THP-1 Cells
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THP-1 cells stably transduced with GFP-actin were maintained in a suspension culture in RPMI media (Gibco, Grand Island, NY, USA), supplemented with 10% FBS (HyClone, Rockford, IL, USA), 1% penicillin streptomycin (Invitrogen, Carlsbad, CA, USA), and 0.05 mM 2-mercaptoethanol. [30 ,36 (link)] For all of the experiments, the cells (13,000 cells/cm2) were plated onto glass coverslips (Ted Pella Inc.) in 35 mm polystyrene petri dishes (BD, Ashland, MA, USA). The THP-1 cells were stimulated with the addition of 1 μg/mL of Phorbol 12-myristate 13-acetate, PMA, for 72 h.
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