The human retinal pigment epithelial cell line (ARPE‐19, CRL‐2302) was obtained from American Type Culture Collection (Manassas, VA, USA). ARPE‐19 cells were grown in DMEM/F‐12 (Lonza, Walkersville, MD, USA) supplemented with 10% foetal bovine serum (FBS; Atlanta Biologicals, Lawrenceville, GA, USA), 1% Glutamax (Lonza) and 1% penicillin–streptomycin (Lonza) at 37°C, 10% CO2. Primary human umbilical vein endothelial cells (HUVECs) were kindly provided by Ms. Case (Center for Excellence in Vascular Biology, Brigham and Women's Hospital, Boston, MA, USA). HUVECs from passages 3 through 6 were cultured in plastic flasks coated with 0.1% gelatin (Sigma‐Aldrich, St. Louis, MO, USA), in EGM‐2 (Lonza) with 20% FBS, SingleQuots supplements (Lonza), 1% Glutamax and 1% penicillin–streptomycin at 37°C, 10% CO2.
Foetal bovine serum fbs
Manufactured by Atlanta Biologicals
Sourced in United States
Foetal bovine serum (FBS) is a nutrient-rich cell culture supplement derived from the blood of bovine fetuses. It is a commonly used component in various cell culture media, providing essential growth factors, proteins, and other biomolecules to support the growth and proliferation of a wide range of cell types in vitro.
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Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Corning (2 mentions)
Dmem f12, by Lonza (1 mentions)
Penicillin streptomycin, by Lonza (1 mentions)
Singlequots supplement, by Lonza (1 mentions)
Gelatin, by Merck Group (1 mentions)
Egm 2, by Lonza (1 mentions)
Arpe 19, by Lonza (1 mentions)
Glutamax, by Lonza (1 mentions)
Dulbecco s modified eagle s medium, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by MP Biomedicals (1 mentions)
Trypsin ethylenediaminetetraacetic acid, by Corning (1 mentions)
Penicillin, by MP Biomedicals (1 mentions)
Streptomycin, by MP Biomedicals (1 mentions)
Amphotericin b, by MP Biomedicals (1 mentions)
Live cell imaging solution, by Thermo Fisher Scientific (1 mentions)
Fluo 4 calcium imaging kit, by Thermo Fisher Scientific (1 mentions)
35 mm dishes, by BD (1 mentions)
L glutamine, by Corning (1 mentions)
Penicillin streptomycin pen strep, by Corning (1 mentions)
Dulbecco s modified eagle medium, by Corning (1 mentions)
9 protocols using foetal bovine serum fbs
1
Culture of Retinal Pigment Epithelial and Endothelial Cells
2
Propagation and Maintenance of NIH3T3, BHK-21, and BMSCs
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NIH3T3 fibroblasts, and BHK-21, baby hamster kidney cells, were propagated in Dulbecco’s modified Eagle’s medium (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% foetal bovine serum (FBS; Atlanta Biologicals, Flowery Branch, GA, USA), 2 mM L-glutamine, 1 mM sodium pyruvate, and 100 U/mL penicillin-streptomycin (HyClone, Logan, UT, USA) followed the literature report.38 (link)BMSCs were propagated in Dulbecco’s modified Eagle’s medium supplemented with 10% FBS, 2 mM L-glutamine, 1 mM sodium pyruvate, 100 U/mL penicillin, 1000 U/mL streptomycin, and 0.25 μg/mL amphotericin B (MP Biomedicals, Irvine, CA, USA).
Cell cultures of NIH3T3, BHK-21, and BMSCs were incubated at 37°C in a CO2 incubator with 5% CO2/95% air. The medium was replaced every 3 days and the cells were trypsinized using 0.25% trypsin-ethylenediaminetetraacetic acid (Corning, Corning, NY, USA).
Cell cultures of NIH3T3, BHK-21, and BMSCs were incubated at 37°C in a CO2 incubator with 5% CO2/95% air. The medium was replaced every 3 days and the cells were trypsinized using 0.25% trypsin-ethylenediaminetetraacetic acid (Corning, Corning, NY, USA).
3
Culturing Cells for Hedgehog Pathway Analysis
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Flp‐In‐3T3 and 293FT cells (Life Technologies) were cultured in Dulbecco's modified Eagle's medium (DMEM) containing high glucose (Thermo Scientific) and supplemented with 10% foetal bovine serum (FBS) (Atlanta Biologicals), 1 mM sodium pyruvate, 2 mM l ‐glutamine, 1× MEM non‐essential amino acids solution, penicillin (40 U/ml) and streptomycin (40 μg/ml), in a humidified atmosphere containing 5% CO2 at 37°C. Experiments in Smo−/− MEFs were performed as previously described 44 . To induce ciliation, cells were grown to confluence in medium containing 10% FBS and then switched to medium containing 0.5% FBS with Hh pathway agonists (SHH‐N and 200 nM SAG) for 24 h (for reporter assays and Western blotting) and 4 h (for Smo immunofluorescence staining). SHH‐N‐conditioned media was generated as previously described 45 .
4
Huh7 Cells Exposed to HCCMF RF EMF
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Huh7 cells – seeded into six-well plates or 35 mm dishes (Falcon; Corning) – before receiving either AM RF EMF treatment. 300,000 cells were left to adhere overnight and cultured in Dulbecco's Modified Eagle's Medium (DMEM; Cellgro) supplemented with heat-inactivated foetal bovine serum (FBS, final concentration 10%; Atlanta Biologicals). Cells were exposed to either HCCMF (hepatocellular carcinoma specific AM RF EMF) or no treatment for 30 min, 1 h, 3 h, or 6 h. 30 min before completion of treatment Fluo-4 stain mix was added directly to each well/dish (Fluo-4 Calcium Imaging Kit; Molecular Probes). Dishes/plate were placed on a rocker at room temperature for 5 min of gentle distribution of Fluo-4 stain. Cells were then placed back into the 37 °C incubator and exposed for 30 min to HCCMF (completing the required time of HCCMF exposure). Following completion of treatment exposure, the dishes/plates were placed at room temperature for 30 min (in the dark). Cells were then washed using 1·5 mL of LCIS (Live Cell Imaging Solution; Molecular Probes) then placed in two millilitres of fresh LCIS and fluorescent intensity was read using a Fluostar fluorescent plate reader (BMG LABTECH) (485 excitation/520 emission). All experiments were performed at least twice with representative experiments shown. Statistics: one-way t-test was utilized.
5
Vero E6 Cell Culture Protocol
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Vero 76 cells, clone E6 (Vero E6), were purchased from ATCC and were grown in DMEM (Dulbecco’s Modified Eagle Medium; Corning) supplemented with 10 % (v/v) foetal bovine serum (FBS; Atlanta Biologicals), 1 % (v/v) l -glutamine (Corning) and 1 % (v/v) penicillin/streptomycin (pen-strep; Corning).
6
Isolation and Culture of Murine Mesenchymal Stem Cells
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Bone marrow‐derived mesenchymal stem cells were harvested from femora and tibiae of young C57BL/6J (3 months) and aged C57BL/6J (16 months) mice. Femora and tibiae were dissected out and cleaned of connective tissue. After clipping both ends of the bones, bone marrow was flushed out from femora and tibiae by a syringe to complete culture media that consisted of a‐MEM (Invitrogen, Carlsbad, CA, USA) supplemented with 20% foetal bovine serum (FBS, Atlanta Biologicals, Atlanta, GA, USA), 100 U/ml penicillin (Invitrogen), 100 μg/ml streptomycin (Invitrogen) and 12 μm l ‐glutamine (Invitrogen). The cell suspension was then plated in 10‐cm culture dishes. The medium was changed every 2–3 days. After digestion with 0.25% trypsin/1 mm ethylenediaminetetraacetic acid (EDTA; Sigma‐Aldrich, St. Louis, MO, USA), cells were passaged at confluence.
7
Culturing HT-22 Hippocampal Cell Line
8
Keratinocyte and Fibroblast Cell Culture
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The normal human epidermal HaCaT keratinocyte cell line and the normal BJ human fibroblast cell line were maintained in DMEM containing 10% foetal bovine serum (FBS; Atlanta Biologicals, Lawrenceville, GA, USA), 100 U/ml of penicillin, 100 mg/ml of streptomycin and Eagle's minimum essential medium (MEM) supplemented with 10% FBS and antibiotics, respectively, at 37°C in a 5% CO2 humidified incubator. UVB irradiation was applied using a bank of 4 Westinghouse F520 lamps (National Biological, Twinsburg, OH, USA) at 6 J/sec./m light in the UVB range. Approximately, 10% of the remaining radiation from the F520 lamp was in the UVA region (320 nm). The UVB exposure chamber was fitted with a Kodak Kodacel K6808 filter to eliminate all wavelengths below 290 nm. UVB radiation was measured using a UVX radiometer (UVX‐31). Transfection was performed with JetPEI (Polyplus‐Transfection, Inc., New York, NY, USA) following the manufacturer's protocol. The cells were cultured for 36–48 hrs and proteins extracted for further analysis. DNA used for CA‐MEK1 and DN‐ERK2 was prepared as described in a previous report 19 .
9
Culturing HEK293T-ACE2 Cell Line
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The Human Embryonic Kidney Cells Expressing Human Angiotensin-Converting Enzyme 2 (HEK293T-ACE2) cell line was obtained from BEI resources (Catalog No. NR-52511, MD, USA), cultured and maintained in DMEM containing 10% foetal bovine serum (FBS) (Atlanta Biologicals, Flowery Branch, GA, USA) and 0.1% penicillin and streptomycin (PS) solution (Life Technologies Corporation, Carlsbad, CA, USA).
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