E. faecalis NBRC 100480 was cultivated and protoplasts were prepared as previously described [23 (link)]. The protoplasts were centrifuged at 7000 rpm for 5 min and resuspended in DMB (5 g/L peptone, 1 g/L yeast extract, 0.1 g/L ferric citrate, 19.45 g/L NaCl, 5.9 g/L MgCl2, 3.24 g/L MgSO4, 1.8 g/L CaCl2, 0.55 g/L KCl, 0.16 g/L NaHCO3, 0.08 g/L KBr, 34 mg/L SrCl2, 22 mg/L H3BO3, 8 mg/L Na2HPO4, 4 mg/L Na2SiO3, 2.4 mg/L NaF, and 1.6 mg/L NH4NO3 [BD, Franklin Lakes, NJ]) containing 300 µg/mL penicillin G. The resulting suspension (5 µL) was diluted with 1 mL of DMB containing 300 µg/mL penicillin G (Wako, Osaka) and incubated at 24 °C.
Penicillin g
Manufactured by Fujifilm
Sourced in Japan, United States
Penicillin G is a laboratory equipment used in the production and cultivation of penicillin, a widely used antibiotic. It serves as a critical tool in the pharmaceutical and biotechnology industries for the growth and isolation of the penicillin-producing microorganisms.
Automatically generated - may contain errors
Lab products found in correlation
Streptomycin, by Fujifilm (35 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (10 mentions)
Streptomycin sulfate, by Fujifilm (8 mentions)
Fetal bovine serum (fbs), by Fujifilm (7 mentions)
Dulbecco modified eagle medium (dmem), by Fujifilm (7 mentions)
Imdm basic medium, by Thermo Fisher Scientific (4 mentions)
Amphotericin b, by Fujifilm (4 mentions)
Fibronectin, by Corning (3 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (3 mentions)
Puromycin, by Merck Group (3 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Na2hpo4, by BD (2 mentions)
Na2sio3, by BD (2 mentions)
Nh4no3, by BD (2 mentions)
B27 supplement, by Thermo Fisher Scientific (2 mentions)
Trypsin edta, by Thermo Fisher Scientific (2 mentions)
Mem basic medium, by Thermo Fisher Scientific (2 mentions)
L glutamine, by Fujifilm (2 mentions)
Trypsin, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Biosera (2 mentions)
52 protocols using penicillin g
1
Cultivation and protoplast preparation of E. faecalis
2
Cultivating E. faecalis Protoplasts
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E. faecalis NBRC 100480 was cultivated and protoplasts were prepared as previously described (Kami et al., 2019 (link)). The protoplasts were centrifuged at 7000 r.p.m. for 5 min and resuspended in Difco Marine Broth 2216 (DMB; 5 g/L peptone, 1 g/L yeast extract, 0.1 g/L ferric citrate, 19.45 g/L NaCl, 5.9 g/L MgCl2, 3.24 g/L MgSO4, 1.8 g/L CaCl2, 0.55 g/L KCl, 0.16 g/L NaHCO3, 0.08 g/L KBr, 34 mg/L SrCl2, 22 mg/L H3BO3, 8 mg/L Na2HPO4, 4 mg/L Na2SiO3, 2.4 mg/L NaF, and 1.6 mg/L NH4NO3 [BD, Franklin Lakes, NJ]) containing 300 µg/mL penicillin G. 2 mL of DMB containing 300 µg/mL of penicillin G (Wako, Osaka) was added to the resulting suspension (10 µL) and incubated at 24 °C.
3
Expansion and Conditioning of Mouse CDCs
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Mouse CDCs were expanded using methods similar to those previously described [23 (link)]. Briefly, atria from mice were minced into small fragments and cultured as “explants” on dishes coated with 15 μg/ml fibronectin (CORNING), by using IMDM basic medium (Gibco), supplemented with 10% fetal bovine serum (HyClone), 100 units/ml penicillin G and 10 μg/ml streptomycin (WAKO, Japan). Stromal-like flat cells and phase-bright round cells emerged from the tissue fragments in 3–5 days and became confluent within 2 weeks. Twice-passaged CDCs were used for the experiments.
Mouse embryonic fibroblasts (MEFs) were purchased from company (EmbryoMax® PMEF-P3, strain CF-1, Millipore, Billerica, MA) and cultured on 0.1% (w/v) gelatin-coated culture dishes in high glucose DMEM medium (Wako, Japan), supplemented with 10% fetal bovine serum, 100 units/ml penicillin G and 10 μg/ml streptomycin.
Totally confluent CDCs or MEFs were changed with fresh medium, and conditioned media were obtained 24 hours later. All cells were cultured in a 5% CO2 incubator at 37°C.
Mouse embryonic fibroblasts (MEFs) were purchased from company (EmbryoMax® PMEF-P3, strain CF-1, Millipore, Billerica, MA) and cultured on 0.1% (w/v) gelatin-coated culture dishes in high glucose DMEM medium (Wako, Japan), supplemented with 10% fetal bovine serum, 100 units/ml penicillin G and 10 μg/ml streptomycin.
Totally confluent CDCs or MEFs were changed with fresh medium, and conditioned media were obtained 24 hours later. All cells were cultured in a 5% CO2 incubator at 37°C.
4
Culturing Laryngeal Cancer Cell Lines
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Laryngeal squamous cell carcinoma cell lines UMSCC‐10A, UMSCC‐10B, UMSCC‐11A, UMSCC‐11B, UMSCC‐12, UMSCC‐13, and UMSCC‐25 were obtained from the Head and Neck Cancer Biology Laboratory at the University of Michigan. Human embryonic kidney (HEK) 293T cells were obtained from the American Type Culture Collection (ATCC). For 2D monolayer culturing, cells were maintained in Dulbecco's Modified Eagle's Medium (DMEM) (Invitrogen), supplemented with 10% fetal bovine serum (FBS) (Gibco), 100 U/mL penicillin G, 100 µg/mL streptomycin sulfate (Wako), and 100 µM MEM nonessential amino acids solution (Wako). For 3D sphere cultures, cells were maintained in DMEM/F‐12 (1:1) medium (Invitrogen), 100 U/mL penicillin G, 100 µg/mL streptomycin sulfate (Wako), 2% B‐27 supplement (Invitrogen), 20 ng/mL epidermal growth factor (EGF) (Sigma), and 20 ng/mL basic fibroblast growth factor (bFGF) (Wako) in ultralow attachment culture dishes (Corning). All cells were maintained in a 5% CO2‐humidified atmosphere at 37°C.
5
Pulmonary Artery Smooth Muscle Cell Culture
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PASMCs (passages 5–10) from normal subjects (Lonza, Basel, Switzerland) and IPAH patients (kindly offered by Prof. Jason X.-J. Yuan) (18 (link), 19 (link)) were cultivated in Medium 199 supplemented with fetal bovine serum (FBS, 10%; Thermo Fisher Scientific, Waltham, MA, USA), D-valine (50 μg/ml; MilliporeSigma, Burlington, MA, USA), endothelial cell growth supplement (20 μg/ml; BD Biosciences, Franklin Lakes, NJ, USA), penicillin G (100 U/ml), and streptomycin (100 μg/ml; Fujifilm Wako Pure Chemical, Osaka, Japan) at 37°C.
6
Caco-2 Cell Proliferation Assay
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Caco-2 cells (ATCC, HTB-37) were cultured in MEM basic medium (Gibco) supplemented with Earle’s salts, L-glutamine and 20% fetal bovine serum. After detachment with 0.25% Trypsin-EDTA (Gibco), the cells were cultured in a 96-well plate at approximately 2 × 104 cells/100 μl/well at 37 °C under 5% CO2 for 24 h. One hundred microliters of 2 μM recombinant Igl, PBST or medium containing 200 units/ml penicillin G and 200 μg/ml streptomycin (Wako, Japan) was added and the cells were incubated for an additional 12 or 24 h under the same conditions. The cells were trypsinized and harvested after 0, 12 or 24 h of incubation and the number of cells per well was counted. The results are shown as the mean of 5 experiments with SD.
7
Cell Culture of Breast Cancer Lines
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The human breast cancer cell lines MDA-MB-231 and MCF-7 were cultured at 37 °C in a 5% CO2 atmosphere in Dulbecco's modified Eagle's medium (DMEM; Nacalai Tesque, Inc.; Kyoto, Japan) supplemented with 10% fetal bovine serum (FBS) and 10,000 units/mL penicillin G, 10,000 μg/mL streptomycin, and 29.2 mg/mL L-glutamine (Wako Pure Chemical Industries, Ltd.; Osaka, Japan). The human mammary epithelial cell line MCF-10A was cultured at 37 °C in a 5% CO2 atmosphere in DMEM/Nutrient Mixture F-12 (Thermo Fisher Scientific Inc.; Waltham, MA, USA) supplemented with 100 ng/mL cholera toxin (Sigma-Aldrich; St. Louis, MO, USA), 20 ng/mL epidermal growth factor (PeproTech Inc.; Rocky Hill, NJ, USA), 0.01 mg/mL insulin (Sigma-Aldrich), 500 ng/mL hydrocortisone (Sigma-Aldrich), and 5% horse serum (Thermo Fisher Scientific Inc.).
8
Cell Line Cultivation and Maintenance
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HEK293T cells and HaCaT cells were obtained from the ATCC and Dr N.E. Fusenig, respectively. These were cultured in DMEM (Sigma Chemical Co., St Louis, MO, USA) supplemented with 10% FBS, penicillin G (100 U/mL), and streptomycin sulfate (0.1 mg/mL; Wako Pure Chemical Industries, Ltd, Osaka, Japan). HaCaT cells stably expressing FLAG‐TSC‐22 were maintained in culture medium supplemented with 1 μg/mL puromycin (Sigma). MGZ5 ES cells were maintained on feeder‐free, gelatin‐coated plates in leukemia inhibitory factor (LIF)‐supplemented medium as described previously.19
9
Expansion of Mouse Cardiac Stem Cells
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Mouse CSCs were expanded using methods similar to those previously described [16 (link)]. Briefly, atrial tissues from mice were minced into small fragments and cultured as “explants” on 6-cm culture dishes coated with 15 μg/ml fibronectin (CORNING). Within 1 week, stromal-like flat cells and phase-bright round cells emerged from the tissue fragments, and they became confluent at approximately 2 weeks. The outgrowth of CSCs was harvested using 0.25% trypsin (Gibco) at 2 weeks, counted using a Nucleo Counter cell-counting device (Chemotetec A/S, Denmark), and then passaged for cell expansion. Twice-passaged CSCs were used for the following experiments as indicated. All cells were cultured in a 5% CO2 incubator at 37°C using IMDM basic medium (Gibco) supplemented with 10% fetal bovine serum (HyClone), 100 units/ml penicillin G and 10 μg/ml streptomycin (WAKO, Japan).
10
HepG2 Cell Culture Protocol
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HepG2 (ATCC number HB-8065) cells were obtained from the ATCC (Baltimore, MD, USA) and cultured in DMEM (Wako, Osaka, Japan) supplemented with 10% fetal bovine serum and antibiotics (100 units/mL penicillin G and 100 μg/mL streptomycin, Wako). The cells were grown at 37°C in 5% CO2 as previously described [21 (link)] and harvested after 1 day of incubation from the last passage.
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