Human osteosarcoma cell lines SaOS-2 were obtained from Dr. Massimo Serra’s Laboratory (IRCCS Istituto Ortopedico Rizzoli, Bologna, Italy) and cultured in Dulbecco’s Modified Eagle Media supplemented with 10% heat-inactivated fetal bovine serum (FBS; Sigma Aldrich, St. Louis, USA), 2 mM L-glutamine, 100 U/ml penicillin, and 100 µg/ml streptomycin (all from MP Biomedicals, Milan, Italy) at 37°C humidified atmosphere with 5% CO2. Human synovial sarcoma cell line SYO-1, bearing the pathogenetic translocation (X;18)(p11.2;q11.2), was obtained from Dr Akira Kawai (National Cancer Center, Tokyo, Japan) and Dr. Aki Yoshida Laboratories (Okayama University, Tokyo, Japan) and cultured in Iscove’s Modified Dulbecco’s Media supplemented with 10% heat-inactivated FBS (Sigma Aldrich), 2 mM L-glutamine, 100 U/ml penicillin, and 100 µg/ml streptomycin (all from MP Biomedicals) at 37°C humidified atmosphere with 5% CO2. The cells were split twice a week. Before cell treatment or usage, SaOS-2 and SYO-1 were removed from culture flasks using 0.05% trypsin-ethylenediamine tetraacetic acid (Life Technologies, Waltham, USA) and washed in PBS. After cell-counting, the cells were resuspended in PBS or cell culture medium.
Streptomycin
Manufactured by MP Biomedicals
Sourced in United States, Italy, Japan, Australia
Streptomycin is a broad-spectrum antibiotic used in laboratory settings. It is effective against a variety of bacterial species, including Gram-positive and Gram-negative bacteria. Streptomycin functions by inhibiting protein synthesis in bacterial cells, leading to cell death.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by MP Biomedicals (57 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (16 mentions)
Amphotericin, by MP Biomedicals (7 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (6 mentions)
Amphotericin b, by MP Biomedicals (4 mentions)
Rpmi 1640 medium, by Lonza (4 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (4 mentions)
Penicillin g, by MP Biomedicals (3 mentions)
Rpmi 1640 media, by Corning (3 mentions)
L glutamine, by Merck Group (3 mentions)
Penicillin, by Merck Group (3 mentions)
Glutamax, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by MP Biomedicals (2 mentions)
H1299, by American Type Culture Collection (2 mentions)
Ampicillin, by MP Biomedicals (2 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (2 mentions)
2 mercaptoethanol, by Nacalai Tesque (2 mentions)
Rpmi 1640 medium, by Merck Group (2 mentions)
Human ab serum, by Gemini Bio (2 mentions)
Magnetic separation column, by Miltenyi Biotec (2 mentions)
74 protocols using streptomycin
1
Culturing Human Osteosarcoma and Synovial Sarcoma Cell Lines
2
Antibiotic Resistance Assessment of Probiotics
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The microdilution protocol was used to assess the resistance of probiotic bacteria to different types of antibiotics [23 (link)] with some modifications according to EFSA Guidance [24 (link)]. The antibiotics ampicillin, chloramphenicol, erythromycin, gentamycin, kanamycin (Sigma Aldrich, St. Louis, MO), and streptomycin (MP Biomedicals, Santa Ana, CA) and the amounts of the active compound were placed into the broth media tube: ampicillin (10 μg), chloramphenicol (30 μg), erythromycin (15 μg), gentamycin (10 μg), kanamycin (30 μg), and streptomycin (10 μg). Cultures were inoculated in MRS broth filtered through a 0.22-μm filter and incubating aerobically at 37°C overnight. Furthermore, antibiotic stock solutions were prepared according to the following formula: where P is potency given by the manufacturer (μg /mg), V is volume required (ml), C is final concentration of solution (multiples of 1000, mg/L), and W is weight of antibiotic (mg) to be dissolved in volume V (mL). The data was expressed after 24 hours as the following formula: logN1 is absorbance of culture 620 nm in MRS broth with different antibiotic types and logN0 is absorbance of culture 620 nm in MRS broth as a control.
3
Isolation and Culture of Rat MSCs and Chondrocytes
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MSCs and chondrocytes from 4 to 5 months old SD-rats were obtained as described previously 27, 28 . MSCs were derived from bone marrow and grown in Dulbecco's modified eagle mediumhigh glucose (DMEM-HG) containing 20% fetal bovine serum (FBS) (SigmaeAldrich, St. Louis, MO, USA), 100 U/ml penicillin and 100 mg/ml streptomycin (MP Biomedicals, Santa Ana, CA, USA).
Cartilage pieces were digested in collagenase D solution (3 mg/ml) (MP Biomedicals, Santa Ana, CA, USA). Isolated chondrocytes were grown in Dulbecco's modified eagle medium-low glucose (DMEM-LG) containing 20% FBS (SigmaeAldrich, St. Louis, MO, USA), 100 U/ml penicillin and 100 mg/ml streptomycin (MP Biomedicals, Santa Ana, CA, USA). Cells at second passage were used in subsequent experiments.
Cartilage pieces were digested in collagenase D solution (3 mg/ml) (MP Biomedicals, Santa Ana, CA, USA). Isolated chondrocytes were grown in Dulbecco's modified eagle medium-low glucose (DMEM-LG) containing 20% FBS (SigmaeAldrich, St. Louis, MO, USA), 100 U/ml penicillin and 100 mg/ml streptomycin (MP Biomedicals, Santa Ana, CA, USA). Cells at second passage were used in subsequent experiments.
4
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).
5
Isolation and Culture of Primary Rat Hepatocytes and LSECs
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The primary hepatocytes and LSECs were harvested from two adult female Lewis rats that weighed between 170 and 200 grams. Animal care and surgical procedures were conducted as per procedures approved by Virginia Polytechnic Institute and State University’s Institutional Animal Care and Use Committee. Hepatocytes were isolated using a two-step collagenase perfusion method as previously described [29 (link)–31 (link), 82 ]. Viability of isolated hepatocytes were ≥ 97% as determined by trypan blue exclusion. Hepatocytes were cultured on type I collagen coated 12-well plates at a density of 500,000 cells per well. Hepatocyte cultures were maintained in high glucose (25 mM) DMEM supplemented with 10% (v/v) heat inactivated fetal bovine serum (FBS), 200 U/mL penicillin, 200 μg/mL streptomycin, 0.5 U/mL insulin (MP Biomedicals, Solon, OH), 4 nM glucagon, and 16 μM hydrocortisone. LSECs were purified from the same isolation using a differential adhesion method [30 (link), 31 (link)] and were cultured on a fibronectin coated flask for 72 hours. LSECs were maintained in endothelial cell medium consisting of Media 199 supplemented with 10% (v/v) heat inactivated FBS, 100 U/mL penicillin, 100 μg/mL streptomycin, 2 mM L-glutamine, and 30 μg/mL endothelial cell growth supplement. Both hepatocytes and LSECs were cultured in their respective media for 72 hours with media exchanges daily.
6
Isolation and Culture of Keloid Fibroblasts
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KFBS were obtained from chest and earlobe keloid tissues of two Japanese
patients. The protocol for tissue collection was approved by the ethics review
board at Kobe University Graduate School. Five or 6 pieces of the minced and
deepithelialized samples were placed on a 100-mm tissue culture dish (Iwaki,
Tokyo, Japan) and immersed in 2 mL Dulbecco's modified Eagle medium (DMEM; Wako
Pure Chemical Industries, Osaka, Japan) supplemented with 10% fetal bovine serum
(FBS; Nichirei, Tokyo, Japan), penicillin (50 U/mL), and streptomycin (50
µg/mL; MP Biomedicals, Illkirch, France). On the following day, 15 mL of
the medium was added to each dish. The culture medium was changed every 2 or 3
days until approximately 80% confluence was reached. The cells were passaged by
incubation at 37°C with 0.05% trypsin and 0.02% EDTA, and plated in culture
dishes.
patients. The protocol for tissue collection was approved by the ethics review
board at Kobe University Graduate School. Five or 6 pieces of the minced and
deepithelialized samples were placed on a 100-mm tissue culture dish (Iwaki,
Tokyo, Japan) and immersed in 2 mL Dulbecco's modified Eagle medium (DMEM; Wako
Pure Chemical Industries, Osaka, Japan) supplemented with 10% fetal bovine serum
(FBS; Nichirei, Tokyo, Japan), penicillin (50 U/mL), and streptomycin (50
µg/mL; MP Biomedicals, Illkirch, France). On the following day, 15 mL of
the medium was added to each dish. The culture medium was changed every 2 or 3
days until approximately 80% confluence was reached. The cells were passaged by
incubation at 37°C with 0.05% trypsin and 0.02% EDTA, and plated in culture
dishes.
7
BEAS-2B Cell Culture and Stimulation
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BEAS-2B cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in Dulbecco's modified Eagle's medium (Invitrogen, Carlsbad, CA, USA) supplemented with 4 mM glutamine (Gibco Invitrogen, Carlsbad, CA, USA), 10% (v/v) fetal calf serum, 100 units/mL penicillin G, and 100 μg/mL streptomycin (MP Biomedicals, Irvine, CA, USA) at 37 °C in a humidified atmosphere with 5% CO2. Reagents for cell stimulation, including the toll-like receptor (TLR) ligands lipoteichoic acid (LTA), polyinosinic:polycytidylic acid (poly I:C), lipopolysaccharide (LPS), and flagellin were purchased commercially (LTA, LPS, and flagellin were purchased from InvivoGen, San Diego, CA, USA; poly I:C was purchased from R&D Systems, Minneapolis, MN, USA). Recombinant human IFN-β was purchased from Abcam (Cambridge, UK). The IFN-β signaling pathway inhibitor, Janus kinase (JAK) inhibitor I, was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The TLR3-neutralizing antibody was purchased from Abcam.
8
Culturing Human Prostate Cancer Cells
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Human prostate cancer cells lines DU145 and LNCaP were cultured in RPMI‐1640 medium (Wako Pure Chemical Industries, Osaka, Japan) supplemented with 10% heat‐inactivated FBS (AusGeneX Pty Ltd, Brisbane, Australia), 100 μg/mL streptomycin (MP Biomedicals, Irvine, CA, USA) and 100 units/mL penicillin (MP Biomedicals) at 37°C in a humidified atmosphere with 5% CO2.
9
Cell Culture of Mouse Fibroblasts and Melanoma
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Mouse fibroblast L929 (NCTC clone 929) and mouse B16-F10 melanoma cell lines were acquired from the Institute of Cytology of the Russian Academy of Sciences (St. Petersburg, Russia). The two cell lines were cultured in DMEM (Dulbecco’s Modified Eagle Medium; Thermo Fisher Scientific, Waltham, MA, USA) with 10% fetal bovine serum (FBS; HyClone, Washington, WA, USA) and 1% antibiotic–antimycotic solution (10 mg/mL streptomycin, 10,000 U/mL penicillin, and 25 μg/mL amphotericin (MP Biomedicals, Santa Ana, CA, USA)) at 37 °C in a humidified atmosphere containing 5% CO2 (standard conditions) and regular passages to maintain exponential growth.
10
Characterization of Human and Murine Lung Cancer Cell Lines
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