Human THP-1 monocytes and RAW 264.7 murine macrophages cells were purchased from Riken Cell Bank. The THP-1 cells were cultured in RPMI supplemented with 10% FBS (Nissui Pharmaceutical Co., Ltd., Tokyo, Japan) at 37 °C in 5% CO2. The THP-1 cells were passaged every 2–4 days at a ratio of 1:5–10. In the experiment, THP-1 cells were differentiated into macrophage-like cells by adding 1 μM PMA (Sigma–Aldrich Corp., St. Louis, MO, USA) to the cells at the time of seeding. The RAW 264.7 cells were cultured in DMEM supplemented with 10% FBS (Nissui Pharmaceutical Co., Ltd.) at 37 °C in 5% CO2. The RAW 264.7 cells were passaged every 2–4 days at a ratio of 1:5–10.
Fetal bovine serum (fbs)
Manufactured by Nissui Pharmaceutical
Sourced in Japan
FBS is a laboratory reagent used as a cell culture supplement. It provides essential nutrients and growth factors to support the in vitro growth and proliferation of various cell types.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Nissui Pharmaceutical (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Dulbecco s modified eagle medium, by Nissui Pharmaceutical (2 mentions)
Fcwf 4, by American Type Culture Collection (2 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Rpmi 1640, by Nissui Pharmaceutical (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium, by Merck Group (1 mentions)
Penicillin, by Nacalai Tesque (1 mentions)
Streptomycin, by Nacalai Tesque (1 mentions)
Fetal bovine serum (fbs), by Fujifilm (1 mentions)
Dulbecco modified eagle medium (dmem), by Nacalai Tesque (1 mentions)
Roswell park memorial institute (rpmi), by Nissui Pharmaceutical (1 mentions)
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
Eagle s minimal essential medium, by Nissui Pharmaceutical (1 mentions)
Afatinib, by Selleck Chemicals (1 mentions)
Osimertinib, by Selleck Chemicals (1 mentions)
Erlotinib, by Selleck Chemicals (1 mentions)
Epoxomicin, by Peptide Institute (1 mentions)
Panc 1, by Nissui Pharmaceutical (1 mentions)
12 protocols using fetal bovine serum (fbs)
1
Monocyte and Macrophage Cell Culture
2
HTLV-1-Infected T Cell Culture
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Normal (HTLV-1-uninfected) CD4+ T cells were obtained from Lonza. The HTLV-1-infected cell line MT-2, ATL-derived cell lines MT-1, TL-Om1 and KOB, and Jurkat cells were cultured in RPMI1640 (Invitrogen) with 10% of fetal bovine serum (FBS; GIBCO) and antibiotics (GIBCO). 293T and 293FT cells were cultured in Dulbecco’s Modified Eagle Medium (Nissui, Japan) with 10% of FBS and antibiotics. All cell lines and primary cultures were maintained at 37 °C with 5% CO2.
3
Feline Coronavirus Propagation and Titration
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fcwf-4 (American Type Culture Collection) was maintained in Dulbecco’s modified Eagle’s medium (Sigma-Aldrich) containing 10 % (v/v) FBS (JRH, Nissui). FCoV (strain 79–1146) was a gift from Dr Tsutomu Hodatsu, Kitasato University, Japan, and was propagated in fcwf-4 cells. Viruses were purified using linear sucrose-gradient ultracentrifugation. Each cell line was infected with FCoV at a m.o.i. of 1. Whole-cell lysates, RNAs and culture fluids were collected 20 h after infection. FCoV titres were expressed as TCID50. The viral stocks were prepared by collecting the virus particles from the culture medium, and TCID50 of each viral preparation was determined by infection of 8 wells of fcwf-4 cells in 96-well plates, in triplicate, with 10-fold serial dilutions of each viral stock.
4
SARS-CoV-2 Virus Propagation and Titration
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All cell cultures were maintained at 37 °C with 5% CO2. Vero E6/TMPRSS2 cells [10 (link)] were obtained from JCRB Cell Bank (Japan) and cultured in Dulbecco’s modified Eagle’s medium (DMEM, Nacalai Tesque, Inc. Kyoto, Japan) supplemented with 10% fetal bovine serum (FBS, Life Technologies), 100 units/ml penicillin, and 100 μg/ml streptomycin (Nacalai Tesque, Inc., Kyoto, Japan).
The SARS-CoV-2 isolate JPN/TY/WK-521 was kindly provided by the National Institute of Infectious Diseases, Japan. It was propagated in a Vero E6/TMPRSS2 cell culture and stored at − 80 °C until use. To determine the infectious titer, Vero E6/TMPRSS2 cells were seeded in 24-well plates at 1.5 × 105 cells/well. On the next day, they were incubated with serial dilutions of virus samples in DMEM without FBS at 37 °C for 2 h. After removal of the supernatant, the cells were cultured in Eagle’s minimum essential medium (EMEM, Nissui Pharmaceutical Co., Ltd., Tokyo, Japan) containing 1% methylcellulose 4000 cp (FUJIFILM, Wako Pure Chemical Corporation, Osaka, Japan) and 2% FBS. Two days after infection, the cells were fixed with formaldehyde, and plaques were visualized by staining with 1% crystal violet. The virus titer was calculated as plaque-forming units/ml.
The SARS-CoV-2 isolate JPN/TY/WK-521 was kindly provided by the National Institute of Infectious Diseases, Japan. It was propagated in a Vero E6/TMPRSS2 cell culture and stored at − 80 °C until use. To determine the infectious titer, Vero E6/TMPRSS2 cells were seeded in 24-well plates at 1.5 × 105 cells/well. On the next day, they were incubated with serial dilutions of virus samples in DMEM without FBS at 37 °C for 2 h. After removal of the supernatant, the cells were cultured in Eagle’s minimum essential medium (EMEM, Nissui Pharmaceutical Co., Ltd., Tokyo, Japan) containing 1% methylcellulose 4000 cp (FUJIFILM, Wako Pure Chemical Corporation, Osaka, Japan) and 2% FBS. Two days after infection, the cells were fixed with formaldehyde, and plaques were visualized by staining with 1% crystal violet. The virus titer was calculated as plaque-forming units/ml.
5
Medaka Cell Line Culture Protocol
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The OLHE-131 cell line was purchased from RIKEN BRC Cell Bank. OLHNI-2, OLKaga-e1, OLHdrR-e3, OLCAB-e21, and OLCAB-e31 [29 (link)] were provided by H. Mitani. All the medaka cell lines were cultured at 30°C in Leibovitz’s L-15 medium (L-15) (Invitrogen, Carlsbad, CA, USA) supplemented with 15% fetal bovine serum (FBS) (Nissui, Tokyo, Japan). At 16 h before transfection, medaka cells were seeded on 12- and 24-well cell culture plates (Sumitomo Bakelite, Tokyo, Japan) at the cell density of 3.2 and 1.6 × 105 cells per well, respectively, which achieved about 80% confluency at the time of transfection.
6
Cell Culture Protocols for BLV, BIV, and BFV
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FLK-BLV cells (which are persistently infected with BLV), CC81 (a feline cell line transformed by mouse sarcoma virus), and CC81-BLU3G and CC81-GREMG cells (derivatives of CC81) [5 (link)] were cultured at 37 °C in 5% CO2 in Dulbecco’s modified Eagle’s Medium (DMEM) (Thermo Fisher, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA). BIV-infected bovine embryonic spleen cells and BFV-infected fetal bovine muscle cells were cultured at 37 °C/5% CO2 in Eagle’s minimal essential medium (Nissui Pharmaceutical Co., Ltd., Tokyo, Japan) supplemented with 10% FBS, as described previously [17 (link), 18 (link)].
7
Characterization of EGFR-Mutant Esophageal Cancer Cell Lines
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The Ba/F3 cell line was maintained in IL‐3 additive RPMI1640 medium (Sigma‐Aldrich, St. Louis, MO, USA) with 10% FBS (Sigma‐Aldrich). Conditioned medium from WEHI‐3 cells was used as a source of IL‐3, as previously described.27 The KYSE270 and KYSE450 cell lines (human esophageal cancer cell lines) were maintained in a 1:1 mixture of RPMI1640 and F12 (Nissui Pharmaceutical, Tokyo, Japan) with 2% FBS according to a previously reported method.30 , 31 , 32 According to the Catalogue of Somatic Mutations in Cancer (COSMIC) database (http://cancer.sanger.ac.uk/cancergenome/projects/cosmic/ ), these cell lines carry uncommon EGFR mutations (KYSE270, L861Q and KYSE450, S768I, respectively). All the cell lines were maintained in a 5% CO2‐humidified atmosphere at 37°C. Erlotinib, afatinib and osimertinib were purchased from Selleck Chemicals (Houston, TX, USA).
8
Pancreatic Cancer Cell Line Cultivation
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Human pancreatic cancer cell lines MIAPaCa-2, AsPC-1, BxPC-3, and PANC-1 were purchased from the American Type Culture Collection (Manassas, VA). Cells were cultured in Dulbecco's modified eagle medium (DMEM) (MIAPaCa-2 and PANC-1) and Roswell Park Memorial Institute medium 1640 (AsPC-1, BxPC-3) purchased from Nissui (Tokyo, Japan) and supplemented with 10% fetal bovine serum (HyClone, Logan, UT). A proteasome inhibitor, epoxomicin, was purchased from Peptide Institute (Ibaraki, Japan).
9
Signaling Pathways in Cell Culture
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Dulbecco's modified Eagle's medium (DMEM), alphamodification of Eagle's medium (aMEM), and fetal bovine serum (FBS) were purchased from Nissui Pharmaceutical Co. Ltd. (Tokyo, Japan), ICN Biomedicals (Aurora, OH) and Nichirei Bioscience Inc. (Tokyo, Japan), respectively. Plastic dishes and multiwell plates were obtained from Greiner Bio-One (Frickenhausen, Germany). All of the inhibitors and actinomycin D were purchased from Merck Millipore (Darmstadt, Germany). Ionomycin and anti-b-actin antibody were obtained from Sigma (St. Louis, MO) and Wako Pure Chemical (Osaka, Japan), respectively. Anti-phospho-extracellular signal-regulated kinase (ERK)1/2, and anti-phospho-p38 MAPK antibody were obtained from Promega (Madison, WI). Anti-ERK1/2, and anti-p38 MAPK antibodies were from Cell Signaling Technology (Beverly, MA). Fluo 4-AM were purchased from Dojindo Laboratories (Kumamoto, Japan).
10
Collagen Synthesis in Lung Fibroblasts
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