Human lung fibroblast cells MRC-5 purchased from the Bioresource Collection and Research Center (BCRC, Food Industry Research and Development Institute, Hsinchu, Taiwan) were cultured in Minimal Essential Medium with Earle’s (MEM) supplemented with 2.2 g/L sodium bicarbonate (NaHCO3), 10% fetal bovine serum (FBS), 1% L-glutamine, 1% sodium pyruvate, 1% non-essential amino acids and 1% penicillin-streptomycin in a humidified incubator at 37 °C with 5% carbon dioxide (CO2). Before treatments, MRC-5 cells were starved in MEM without FBS for 24 h. Afterwards, MRC-5 cells were pre-treated with or without Q3G (0.125 μM or 0.25 μM), Dex (10 μM) [26 (link)], or solvent dimethyl sulfoxide (DMSO, 0.125%), served as solvent control, for 6 h, and then co-incubated with LPS (10 μg/mL) for another 24 h. For the preparations of LPS, Q3G and Dex, by dissolving the powder of LPS from Sigma–Aldrich (St Louis, MO, USA) in sterilized water, LPS at the stock concentration (10 mg/mL) was prepared. Q3G (CAS no. 22688-79-5; purity >98.0%) was obtained from ChemFaces (Wuhan, Hubei, China) and prepared the stock concentration (200 μM) by dissolving the powder in DMSO. Dex, purchased from Sigma–Aldrich (St Louis, MO, USA), was prepared the stock concentration (5 mM) by dissolving the powder in 50% (v/v) ethanol.
Mrc 5
Manufactured by RIKEN BioResource Center
Sourced in Japan
The MRC-5 is a type of cell culture that is derived from normal human lung fibroblast cells. It is a widely used cell line in biomedical research and vaccine production.
Automatically generated - may contain errors
Lab products found in correlation
Nci h460, by RIKEN BioResource Center (2 mentions)
Nci h520, by RIKEN BioResource Center (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Mrc 5, by Merck Group (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Mkn45, by RIKEN BioResource Center (1 mentions)
Nugc 4, by RIKEN BioResource Center (1 mentions)
Katoiii, by RIKEN BioResource Center (1 mentions)
Minimum essential medium (mem), by Thermo Fisher Scientific (1 mentions)
Mrc 5, by Thermo Fisher Scientific (1 mentions)
Z vad fmk, by Santa Cruz Biotechnology (1 mentions)
Coronarin d, by ChemFaces (1 mentions)
Jnk inhibitor sp600125, by Santa Cruz Biotechnology (1 mentions)
Penicillin, by Sartorius (1 mentions)
Streptomycin, by Sartorius (1 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
Raw264, by Thermo Fisher Scientific (1 mentions)
Raw 264, by RIKEN BioResource Center (1 mentions)
14 protocols using mrc 5
1
Suppression of LPS-induced inflammation in human lung fibroblasts
2
Cell Culture of Lung Cell Lines
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Human lung carcinoma cell line A-549 (BCRC 60074), H661 (BCRC 60125) and human lung normal fibrobast MRC-5 (BCRC 60023) were purchased from Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan). A-549 and MRC-5 cells were grown in DMEM with 10% fetal bovine serum, 1% penicillin/streptomycin, and supplemented with 2 mM l -glutamine. H661 cells were grown in RPMI-1640 medium supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin, and 2 mM l -glutamine. Cells were subcultured every two days and cultivated in a humidified incubator at 37 °C with 5% CO2 and 95% air.
3
Gastric Cancer Cell Lines TP53 Profiling
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Seven gastric cancer cell lines were used: four cell lines with wt TP53 (MKN-45, NUGC-4, STKM-2, SNU-1), two cell lines with mutant-type (mt) TP53 (NUGC-3, STKM-1), and one cell line with null TP53 (KatoIII) [19] (link)–[21] (link). Cell lines with wt TP53 (MCF-7 breast cancer, HEK293 human embryonic kidney cells) and MRC-5 normal human fibroblasts were included as controls in this study. MKN-45, NUGC-4, KatoIII, and MRC-5 cell lines were obtained from RIKEN BRC Cell Bank (Tsukuba, Japan). SNU-1 and MCF-7 cell lines were purchased from the American Type Culture Collection (Rockville, MD). NUGC-3 and HEK293 cell lines were obtained from Health Science Research Resources Bank (Osaka, Japan). STKM-1 and STKM-2 cell lines were kindly provided by Dr. Shunsuke Yanoma (Yokohama City University, School of Medicine, Japan).
4
Osteosarcoma Cell Line Treatment
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Human HOS and MG-63 osteosarcoma cells, and human fibroblast cell line MRC-5 were purchased from the Bioresource Collection and Research Center (Hsinchu, Taiwan). MRC-5, HOS, and MG-63 cells were maintained in Minimum Essential Medium (#11095-080; Gibco, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (Gibco), 1% penicillin/streptomycin (Gibco). JNK inhibitor (SP600125) and z-VAD-FMK were purchased from Santa Cruz Biotechnology. Coronarin D (purity > 95%) was purchased from ChemFaces (Wuhan, Hubei, China).
5
Cell Culture Protocols for Lung Cancer Research
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Human lung adenocarcinoma A549 cells, murine LLC1 cells, human lung fibroblast MRC-5, and murine macrophages Raw264.7 were acquired from the Bioresource Collection and Research Center (BCRC; Hsinchu, Taiwan). Human lung adenocarcinoma CL1-5 cells were obtained from Dr. P.-C. Yang of National Taiwan University. The lung cancer cell lines were cultured as previously described [28 (link),29 (link)]. The MRC-5 cells were cultured in minimum essential medium supplemented with 10% heat-inactivated fetal bovine serum (FBS; VWR Life Science Seradigm, Radnor, PA, USA), 2 mM L-glutamine, 0.1 mM nonessential amino acids, 1.5 g/L sodium bicarbonate, and 1.0 mM sodium pyruvate at 37 °C in an environment of 95% air and 5% CO2. The RAW 264.7 cells were kept in Dulbecco’s modified Eagle medium (GIBCO-Life Technologies) that was supplemented with 3.7 g/L NaHCO3, 5% heat-inactivated FBS, and 100 units/mL each of penicillin and streptomycin (Biological Industries, Cromwell, CT, USA).
6
Cell Culture of Various Cell Lines
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NIH/3T3 cells were provided by C. Sherr (St. Jude Children Hospital, Memphis, TN, USA), and U-2 OS, HT1080, and HEK293 cells were provided by M. Ohtsubo (Beppu University, Oita, Japan). MRC-5 were purchased from RIKEN BioResource Center (Tsukuba, Japan). Cell were grown at 37 °C in Dulbecco’s modified Eagle medium (DMEM) containing 10% tetracycline-free fetal bovine serum (Takara bio Inc., Kusatsu, Japan) with 5% CO2.
7
Culture and Maintenance of NSCLC Cell Lines
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The human non-small cell lung cancer (NSCLC) cell line H1299 was obtained from the American Type Culture Collection (Manassas, VA, USA). The NSCLC cell line A549 and normal lung fibroblast cell line MRC-5 were obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan). All cells were cultured in accordance with suppliers’ recommendations. H1299 cells were cultured in Roswell Park Memorial Institute 1640 medium supplemented with 10% fetal bovine serum, 100 U/mL penicillin, and 100 μg/mL streptomycin. A549 cells were cultured in Ham’s F-12 nutrient mixture supplemented with the above-mentioned supplements, whereas MRC-5 cells were cultured in Eagle’s minimum essential medium with the same supplements. Cells were incubated in an incubator under 5% CO2 in air at 37°C.
8
SARS-CoV-2 Strain Propagation and Characterization
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BHK-21 cells and Calu-3 cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Gibco) supplemented with 10% Fetal Bovine Serum (FBS) and 1 × penicillin/streptomycin solution. Vero E6 cells were maintained in High glucose DMEM (GeneDireX) supplemented with 10% FBS. Normal human lung WI-38 VA-13 subline 2RA and MRC-5 cells were purchased from the Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan). Cells were cultured in Minimum essential medium (Eagle, Gibco) supplemented with 10% FBS, 2 mM L-glutamine, 1.5 g/L sodium bicarbonate, 0.1 mM non-essential amino acids, and 1.0 mM sodium pyruvate at 37°C under a mixture of 95% air and 5% CO2. The SARS-CoV-2 strain 3586 (TSGH_15 GISAID accession number EPI_ISL_436100) was isolated at the Institute of Preventive Medicine, National Defense Medical Center and amplified in Vero E6 cells. The viral titer was determined using a plaque assay. SARS-CoV-2 was handled in a BSL-3 laboratory.
9
Lung Cell Line Response to THSG
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Human lung fibroblast cell line MRC-5 was obtained from the Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan). Human lung alveolar epithelial cell line A549 was obtained from the American Type Culture Collection (Manassas, VA, United States). MRC-5 and A549 cells were cultured in MEM and DMEM respectively, supplemented with 10% FBS, 1% sodium pyruvate, and 1% antibiotic-antimycotic solution. Cells were incubated in a humidified chamber at 37°C with 95% air and 5% CO2 environment. In this study, the MRC-5 cells were pretreated with various concentrations of THSG (50 and 100 μg/ml) for 24 h. Afterward, exogenous 2.5 ng/ml of TGF-β1 was administered for 1, 6, and 24 h. Additionally, the A549 cells were pretreated with 50 and 100 μg/ml of THSG for 2 h, and then stimulated with 5 ng/ml of TGF-β1 for 30 min, 1, 3, and 48 h.
10
Cell Culture Conditions for Cell Lines
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Human embryonic kidney 293 cells (HEK293T) were purchased from Thermo Fisher Scientific (Waltham, MA, USA); two human fetal lung fibroblast cells (HFL-1 and MRC-5) and human breast adenocarcinoma cells (MCF-7) were purchased from Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan). The H1299 human non-small cell lung cancer cell line was acquired from the American Type Culture Collection (ATCC, Manassas, VA, USA). The HEK293T, HFL-1, MRC-5, and MCF-7 cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (HyCloneTM, Cytiva, Marlborough, MA, USA) containing 10% fetal bovine serum (FBS; Sigma, St Louis, MO, USA) and 1% penicillin/streptomycin; the H1299 cell was cultured in RPMI (HyCloneTM, Cytiva, Marlborough, MA, USA) containing 10% FBS and 1% penicillin/streptomycin. All cell lines were incubated at 37 °C in a humidified incubator containing 5% CO2.
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