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Streptomycin is a broad-spectrum antibiotic produced by the actinobacterium Streptomyces griseus. It is a crystalline compound used in the treatment of various bacterial infections.

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299 protocols using streptomycin

1

Culturing Colorectal Cancer and Normal Intestinal Cells

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Tumor intestinal epithelial cells HCT-116 (ATCC® CCL-247™) and HT-29 (ATCC® HTB-38™), derived from human colorectal adenocarcinoma, were cultured in DMEM 10% FBS, 10 mM Hepes, 100 U/mL of penicillin, and 100 µg/mL of streptomycin (Sigma, Darmstadt, Germany). Tumor intestinal epithelial cells DLD1 (ATCC® CCL-221™), derived from human colorectal adenocarcinoma, were cultured in RPMI1640 10% FBS, 10 mM Hepes, 100 U/mL of penicillin, and of 100 µg/mL streptomycin. Normal human intestinal cells CCD841 CoN (ATCC® CRL-1790™) were cultured in EMEM 10% FBS, 10 mM Hepes, 100 U/mL of penicillin, and 100 µg/mL of streptomycin. Cells were cultured at 37 °C in a humidified incubator.
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Ovarian Cancer Cell Lines and Nude Mouse Model

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The human ovarian cancer cell lines SK-OV-3 cells (HTB-77, ATCC, Manassas, VA, USA ) and OVK18 cells (TKG 0323, Cell Bank, Tohoku University, Sendai, Japan) were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) (Thermo Fisher Scientific, Waltham, MA, USA), containing 100 U/mL penicillin (Nacalai tesque, Kyoto, Japan), and 100 μg/mL streptomycin and OVCAR-3 (HTB-161, ATCC, Manassas, VA, USA) were cultured in RPMI 1640 medium supplemented with 10% FBS containing 100 U/mL penicillin, and 100 μg/mL streptomycin.
Four-week-old female BALB/c nude mice from Charles River (Kanagawa, Japan) were bred at 23 °C and fed with sterilized food and water during the experiments. All animal experimental protocols were reviewed and approved by the ethics committee (Animal Care and Use Committee) of Okayama University under the project identification code IDs OKU-2016078 (Date of approval: 1 April 2016).
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3

Cultivation of HUH7 and Vero Cell Lines

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All cell lines were maintained in a humidified 5% CO2 environment at 37 °C. HUH7 and HUH7.5 cells (Human hepatocyte derived cells from carcinoma, Apath LLC, new York, NY, USA) were cultured in DMEM (4500 mg/L “high glucose,” + L-glutamine, + 25 mM HEPES, - Sodium Pyruvate; Life Technologies 12430-062) supplemented with fetal bovine serum (FBS, Gibco; 10% FBS was used for cytotoxicity assays and 2% was used in antiviral plaque assays), penicillin (100 I.U./mL), and streptomycin (100 μg/mL, ATCC) and 1% non-essential amino acids (NEAA; Sigma Aldrich M7145-100ML) at a density of 2×105 – 2×106 cells/mL. Vero cells (ATCC, CCL-81) were cultured in EMEM media (ATCC) supplemented with FBS (10% FBS was used for cytotoxicity assays and 2% was used in antiviral plaque assays), penicillin (100 I.U./mL), and streptomycin (100 μg/mL) at a density of 2×105 – 2×106 cells/mL. HUH7 Cells were verified by short tandem repeat (STR) profiling by Creative Bioarray.
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4

NSCLC Cell Line Culturing Protocol

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NSCLC cell lines, A549 (ATCC® CRL-185™) and H2170 (ATCC® CRL-5928™) were cultured in RPMI-1640 (Invitrogen, Carlsbad, CA, USA) medium containing 10% fetal bovine serum (FBS), 100 IU/ml penicillin and 100 µg/ml streptomycin (all purchased from ATCC) and grown at 37°C in a humidified 5% CO2 atmosphere. Human lung fibroblast (IMR-90) cells were cultured in MEM-α (1x)-Glutamax medium containing 10% FBS, 100 IU/ml penicillin and 100 µg/ml streptomycin (ATCC). Cells were maintained in T75 tissue culture flasks, and the medium was changed three times a week. Confluent cells were harvested by washing in phosphate-buffered saline (PBS) followed by trypsinisation (0.25% in EDTA) for subculturing. All of the cell lines were purchased from ATCC, and culture reagents were purchased from Gibco-Life Technologies (Grand Island, NY, USA) unless otherwise stated.
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5

Differentiation and Infection of U937 Cells

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The human pro-monocytic myeloid leukemia cell line U937 (ATCC® CRL-1593.2, USA) was maintained in RPMI containing 10% FCS, penicillin (100 U/mL), and streptomycin (100 μg/mL) (Thermo Fisher Scientific, USA). For each experiment, U937 cells were differentiated into macrophages by incubation with 10 ng/mL PMA (Sigma-Aldrich, USA) for 48 h; then, the supernatant was removed, and fresh medium was added. Macrophage differentiation was assessed by RT-qPCR of CD14 (Supplementary Figure 1). After 48 h in fresh medium, U937 cells were treated for each experiment. T. cruzi incubation was always at a multiplicity of infection (MOI) of 1. This ratio was selected because it assures a 1:1 cell contact without jeopardizing cell viability (Supplementary Figure 2). Vero cells (ATCC® CCL-81, USA) were cultured in RPMI containing 5% FCS, penicillin (100 U/mL), and streptomycin (100 μg/mL). Vero cells were infected with T. cruzi trypomastigotes (Dm28 strain) at an MOI of 5 for parasite amplification. The HEK293T (ATCC® CRL-3216, USA) cell line was cultured in DMEM containing 10% FCS, penicillin (100 U/mL) and streptomycin (100 μg/mL). HEK293T cells were cultured for lentivirus production. All cells were grown at 37°C in 5% CO2 in a humidified atmosphere.
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HCoV-OC43 Culture Protocol

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HCT‐8 epithelial cells (ECACC 90032006) were cultured at 37°C with 5% CO2 in RPMI‐1640 (Lonza, Basel, Switzerland) culture medium supplemented with 10% fetal bovine serum (FBS; HyClone, Logan, UT), 100 IU ml−1 penicillin and 100 μg ml−1 streptomycin (Lonza, Basel, Switzerland). BHK‐21 fibroblast cells (clone 13; ECACC 85011433) were cultured in Dulbecco's Modified Eagle Medium (DMEM; Lonza, Basel, Switzerland) supplemented with 10% FBS, 100 IU ml−1 penicillin and 100 μg ml−1 streptomycin at 37°C with 5% CO2.
Human coronavirus HCoV‐OC43 (ATCC VR‐1558) was cultured in HCT‐8 epithelial cells in RPMI‐1640 with 5% FBS, 100 IU ml−1 penicillin and 100 μg ml−1 streptomycin for 7 days at 33°C. HCoV‐OC43 was harvested by aspirating the RPMI‐1640 culture media and centrifuging (3000g, 4 min) to remove cell debris. The virus stocks were stored at −80°C before use. All experiments were performed within a Biosafety Level 2 laboratory.
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7

Cell Culture Protocols for Viral Studies

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Human astroglial SVG-A derived cells (a kind gift from Walter J. Atwood, Brown University, Providence, RI) were grown at 37 °C and 5% CO2 in Minimum Essential Medium (MEM) (10-010-CV; Corning) supplemented with 10% heat-inactivated fetal bovine serum (FBS, S11150; Atlanta Biologicals), penicillin, and streptomycin (1406-05-9; VWR International). African Green Monkey kidney epithelial MA104 cells (a kind gift from Siyuan Ding, Washington University in St. Louis, St. Louis, MO) were grown at 37 °C and 5% CO2 in Medium 199 supplemented with 10% heat-inactivated FBS. Vero C1008 (Vero 76, clone E6, Vero E6) [American Type Culture Collection (ATCC) CRL-1586] cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% FBS, and penicillin and streptomycin. Vero CCL-81 (ATCC CCL-81) cells were maintained in DMEM supplemented with 10% FBS, 10 mM Hepes pH 7.4, 1% Glutamax, and penicillin/streptomycin.
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Culture of Renal Adenocarcinoma Cell Lines

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The human renal adenocarcinoma cell line 769-P was obtained from ATCC; the cells were maintained in RPMI 1640 medium supplemented with 10% FBS, penicillin (100 U/ml) and streptomycin (100 μg/ml) (Gibco Life Technologies). The human renal adenocarcinoma cell lines ACHN and A498 were purchased from ATCC; the cells were maintained in minimal essential medium (MEM) supplemented with 10% FBS, penicillin (100 U/ml) and streptomycin (100 μg/ml). The human renal adenocarcinoma cell lines Caki-1 and Caki-2 were purchased from ATCC; the cells were maintained in McCoy's 5A medium supplemented with 10% FBS, penicillin (100 U/ml) and streptomycin (100 μg/ml). All cells were incubated in 5% CO2 at 37 °C.
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9

Cell Line Maintenance under Controlled Conditions

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All cell lines were maintained in a humidified 5% CO2 environment at 37 °C in tissue culture flasks (Corning) under normoxic conditions. Adherent cells were dissociated using either Trypsin-EDTA solution (0.25%, Gibco) or TrypLE Express solution (Invitrogen). HL-60, CCRF-CEM, U-87 MG, GBM6, DU-145, and NCI/ADR-RES cells were cultured as described previously.58 (link), 64 , 80 (link) MCF-7 cells (ATCC, HTB-22) were cultured in MEM media (Cellgro) supplemented with 10% FBS (Gibco), bovine insulin (0.01 mg/mL, Sigma), penicillin (100 I.U./mL, ATCC), and streptomycin (100 µg/mL, ATCC). TEX cells73 (link), 74 (link) were cultured in IMDM containing L-glutamine (Cellgro) supplemented with 15% FBS (Gibco), Stem Cell Factor (20 ng/mL, PeproTech), Interleukin-3 (2 ng/mL, PeproTech), penicillin (100 I.U./mL, ATCC), and streptomycin (100 µg/mL, ATCC).
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10

Cell Culture Conditions for Breast Cancer

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MDA-MB-231 and MCF-7 cells were cultured in 37°C incubator with 5% CO2 and maintained in DMEM (Lonza, Walkersville, MD, USA) containing 5% fetal bovine serum (ATCC, Rockville, MD, USA), 100 IU/ml of penicillin and 100 μg/ml streptomycin (ATCC, Rockville, MD, USA). SKBR3 and BT474 cells were cultured in 37°C incubator with 5% CO2 and maintained in RPMI (Lonza, Walkersville, MD, USA) containing 5% fetal bovine serum (ATCC, Rockville, MD, USA), 100 IU/ml of penicillin and 100 μg/ml streptomycin (ATCC, Rockville, MD, USA).
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