gentamicin sulfate, by Thermo Fisher Scientific - Product details

1

Cell Culture Protocol for CHO-K1, HEK293T, and BHK-21 Cells

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CHO-K1 cells were cultured in adherent conditions in chemically defined medium DMEM-F12 (Gibco), supplemented with 5% (v/v) of fetal bovine serum (FBS) (PAA Laboratories), 200 mM L-glutamine (Sigma-Aldrich), 2.441 g/L NaHCO3 (Gibco) and 0.05 mg/mL gentamicin sulfate (Gibco). HEK293T cells were maintained in 13.37 g/L D MEM medium (Gibco), supplemented with 10% (v/v) of FBS (PAA Laboratories), 1.5 g/L NaHCO3 (Gibco), 0.05 g/L gentamicin sulfate (Gibco) and 0.11 g/L sodium pyruvate. BHK-21 cells were cultured in MEM medium (Gibco), supplemented with 10% (v/v) FBS (PAA Laboratories), 2.2 g/L NaHCO3 (Gibco) and 0.05 g/L gentamicin sulfate (Gibco). Cells lines were cultured in a humidified atmosphere at 37 °C and 5% CO2.

Tossolini I., Gugliotta A., López Díaz F., Kratje R, & Prieto C. (2022). Screening of CHO-K1 endogenous promoters for expressing recombinant proteins in mammalian cell cultures. Plasmid, 119-120.

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Cell Culture Protocol for CHO-K1, HEK293T, and BHK-21 Cells

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CHO-K1 cells were cultured in adherent conditions in chemically defined medium DMEM-F12 (Gibco), supplemented with 5% (v/v) of fetal bovine serum (FBS) (PAA Laboratories), 200 mM L-glutamine (Sigma-Aldrich), 2.441 g/L NaHCO3 (Gibco) and 0.05 mg/mL gentamicin sulfate (Gibco). HEK293T cells were maintained in 13.37 g/L D MEM medium (Gibco), supplemented with 10% (v/v) of FBS (PAA Laboratories), 1.5 g/L NaHCO3 (Gibco), 0.05 g/L gentamicin sulfate (Gibco) and 0.11 g/L sodium pyruvate. BHK-21 cells were cultured in MEM medium (Gibco), supplemented with 10% (v/v) FBS (PAA Laboratories), 2.2 g/L NaHCO3 (Gibco) and 0.05 g/L gentamicin sulfate (Gibco). Cells lines were cultured in a humidified atmosphere at 37 °C and 5% CO2.

Tossolini I., Gugliotta A., Díaz F.L., Kratje R., & Prieto C. (2021). Screening of CHO-K1 endogenous promoters for expressing recombinant proteins in mammalian cell cultures.

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Characterization of Mouse Tumor Cell Lines

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TS/A and TUBO mouse mammary carcinoma cell lines were derived from a BALB/c background and were gifts from Dr. Patrizia Nanni (University of Bologna, Bologna, Italy)^{34 (link),35 (link)} and were grown in Dulbecco’s modified Eagle medium (DMEM, BioSource, Inc., Gaithersburg, MD) with 10% fetal bovine serum (FBS; Gemini, Calabassas, CA) and 10 μg/mL gentamicin sulfate (BioSource). Human embryonic kidney (HEK-293) cells and murine TRAMP-C2 prostate cancer cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA) and were grown in DMEM with 10% FBS and 10 μg/mL gentamicin sulfate (BioSource). MC38 murine colon cancer cells were grown in DMEM with 10% FBS and 10 μg/mL gentamicin sulfate (BioSource). TRAMP-C2 and MC38 cells were derived on a C57Bl/6 background. CTLL-2 cytotoxic T cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA) and were cultured in RPMI 1640 supplemented with 10% fetal bovine serum (FBS) and 200 U/ml IL-2.

Morris J.C., Ramlogan-Steel C.A., Yu P., Black B.A., Mannan P., Allison J.P., Waldmann T.A, & Steel J.C. (2014). Vaccination with Tumor Cells Expressing IL-15 and IL-15Rα Inhibit Murine Breast and Prostate Cancer. Gene therapy, 21(4), 393-401.

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4

Characterization of Mouse Tumor Cell Lines

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TS/A and TUBO mouse mammary carcinoma cell lines were derived from a BALB/c background and were gifts from Dr. Patrizia Nanni (University of Bologna, Bologna, Italy)^{34 (link),35 (link)} and were grown in Dulbecco’s modified Eagle medium (DMEM, BioSource, Inc., Gaithersburg, MD) with 10% fetal bovine serum (FBS; Gemini, Calabassas, CA) and 10 μg/mL gentamicin sulfate (BioSource). Human embryonic kidney (HEK-293) cells and murine TRAMP-C2 prostate cancer cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA) and were grown in DMEM with 10% FBS and 10 μg/mL gentamicin sulfate (BioSource). MC38 murine colon cancer cells were grown in DMEM with 10% FBS and 10 μg/mL gentamicin sulfate (BioSource). TRAMP-C2 and MC38 cells were derived on a C57Bl/6 background. CTLL-2 cytotoxic T cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA) and were cultured in RPMI 1640 supplemented with 10% fetal bovine serum (FBS) and 200 U/ml IL-2.

Morris J.C., Ramlogan-Steel C.A., Yu P., Black B.A., Mannan P., Allison J.P., Waldmann T.A, & Steel J.C. (2014). Vaccination with Tumor Cells Expressing IL-15 and IL-15Rα Inhibit Murine Breast and Prostate Cancer. Gene therapy, 21(4), 393-401.

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5

Culturing Breast, Oral Squamous Cell Lines

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MCF-7 breast cancer cells were grown in high glucose DMEM (Invitrogen, Carlsbad, CA) supplemented with 10% FBS (Invitrogen), 1% gentamicin sulfate (Invitrogen), and 10 μg/mL recombinant human insulin (Santa Cruz Biotechnology). MDA-MB-231 cells were cultivated in RPMI 1640 (Invitrogen) with 10% FBS and 1% gentamicin sulfate. SCC15 oral squamous cell carcinoma was grown in high glucose DMEM supplemented with 10% FBS and 1% gentamicin sulfate. All cell lines were grown at 37°C in humidified incubators with 5% CO₂.

Zelasko-Leon D.C., Fuentes C.M, & Messersmith P.B. (2015). MUC1-Targeted Cancer Cell Photothermal Ablation Using Bioinspired Gold Nanorods. PLoS ONE, 10(7), e0128756.

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Cell Culture and Palbociclib Sensitivity

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TKO MEFs were originally provided by Julian Sage [17 (link)] and the A549 and SK-MES-1 lung cancer cells were obtained from ATCC. MEFs and A549 cells were cultured in DMEM (Gibco) supplemented with 10% fetal bovine serum (Hyclone) and 50 μg/ml gentamicin sulfate (Gibco). SK-MES-1 cells were cultured in RPMI-1640 (Gibco) supplemented with 10% fetal bovine serum (Hyclone) and 50 μg/ml gentamicin sulfate (Gibco). Palbociclib (PD0332991) was purchased from Selleck Chemicals. Cell number and viability was determined by trypan blue exclusion and enumeration using a hemocytometer.

Kruer T.L., Dougherty S.M., Reynolds L., Long E., de Silva T., Lockwood W.W, & Clem B.F. (2016). Expression of the lncRNA Maternally Expressed Gene 3 (MEG3) Contributes to the Control of Lung Cancer Cell Proliferation by the Rb Pathway. PLoS ONE, 11(11), e0166363.

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7

Isolation of Lymphocytes from Primate Samples

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Peripheral blood mononuclear cells were isolated from pigtail macaque blood by Histopaque-1077 (Sigma-Aldrich) density gradient in Accupsin conical tubes (Sigma-Aldrich). Red blood cells were lysed using ACK lysis buffer (ThermoFisher) and then resuspended in RPMI-1640 media (Invitrogen) supplemented with 10% Fetal Bovine Serum (FBS) (VWR), 50 µg/ml Gentamicin sulfate (ThermoFisher), and 5 mg/ml of Penicillin–Streptomycin–Glutamine (Pen–Strep–Glut) (ThermoFisher). Intraepithelial and lamina propria lymphocytes of gut and vaginal mucosa biopsies were isolated by straining through a 70-micron filter (Corning) following a 1 hour (h) enzymatic digestion at 37 °C with 40 µg/ml Liberase TM (Sigma-Aldrich) and 80 µg/ml DNase (Sigma-Aldrich) in RPMI-1640 media supplemented with 5 mg/ml of Pen–Strep–Glut (ThermoFisher). Lymphocytes from whole lymph node biopsies were isolated by directly straining through a 70-micron filter in RPMI-1640 media supplemented with 10% FBS (VWR) and 5 ml of 1X Pen–Strep–Glut (ThermoFisher), and 50 μg/ml Gentamicin sulfate (ThermoFisher).

O’Connor M.A., Tisoncik-Go J., Lewis T.B., Miller C.J., Bratt D., Moats C.R., Edlefsen P.T., Smedley J., Klatt N.R., Gale M J.r, & Fuller D.H. (2018). Early cellular innate immune responses drive Zika viral persistence and tissue tropism in pigtail macaques. Nature Communications, 9, 3371.

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8

Cell Culture Conditions for Various Cell Lines

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SKW3 T cells (DSMZ) were cultured in RPMI-1640+GluMax (Thermo Fisher Scientific) complemented with 10% fetal bovine serum (FBS, Sigma-Aldrich), 10 mM HEPES and 50 U/mL Pen-Strep (Thermo Fisher Scientific) at 37 °C and 5% CO₂.
LentiX cells and 293T cells were cultured in DMEM (Thermo Fisher Scientific) supplemented with 10% FBS, 2 mM L-Glutamine, 10 mM HEPES and 50 U/mL Pen-Strep (Thermo Fisher Scientific) at 37 °C and 5% CO₂.
KG-1 cells (ATCC) were cultured in IMDM (Thermo Fisher Scientific) supplemented with 10% FBS and 50 U/mL Pen-Strep (Thermo Fisher Scientific) at 37 °C and 5% CO₂.
SF9 cells were cultured in SF900-III media (Thermo Fisher) supplemented with 10% FBS and 10 mg/mL gentamicin sulfate (Thermo Fisher) at 27 °C and atmospheric CO₂.
Hi5 cells were grown in insect cell culture medium (Expression Systems) supplemented with 10 mg/mL gentamicin sulfate (Thermo Fisher) at 27 °C and atmospheric CO₂.
Jurkat cell lines were cultured in RPMI 1640 supplemented with 10% FBS, 2 mM L-Glutamine, 50 U/mL Penicillin, 50 μg/mL Streptomycin, and 50 μM β-mercaptoethanol at 37 °C and 5% CO₂.
HEK293T cell line was cultured in DMEM supplemented with 10% FBS, 2 mM L-Glutamine, and 18 mM HEPES at 37 °C and 5% CO₂.

Zhao X., Kolawole E.M., Chan W., Feng Y., Yang X., Gee M., Jude K.M., Sibener L.V., Fordyce P.M., Germain R.N., Evavold B.D, & Garcia K.C. (2022). Tuning T cell receptor sensitivity through catch bond engineering. Science (New York, N.Y.), 376(6589), eabl5282.

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9

MAYV Virus Propagation in Cell Lines

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MAYV (VR-1277, genotype D) was obtained from the American Type Culture Collection (Manassas, VA, USA). Baby hamster kidney (BHK-21) and African green monkey kidney (Vero) cells were cultured as monolayers in 25-cm² flasks (TPP, Trasadingen, Switzerland) at 37 °C in a humidified atmosphere with 5% CO₂ in DMEM (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (Cultilab, Campinas, SP, Brazil) and 5 µg/mL gentamicin sulfate (Invitrogen, Carlsbad, CA, USA).

Carvalho C.A., Silva J.L., Oliveira A.C, & Gomes A.M. (2017). On the entry of an emerging arbovirus into host cells: Mayaro virus takes the highway to the cytoplasm through fusion with early endosomes and caveolae-derived vesicles. PeerJ, 5, e3245.

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10

Cellular Signaling Pathway Analysis

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DMEM/Ham's nutrient mixture F-12, diethylstilbestrol (DES), atRA, and H-89 dihydrochloride hydrate were purchased from Sigma-Aldrich, Inc. (St. Louis, MO, USA). Gentamicin sulfate and fungizone were purchased from Invitrogen Corp. (Carlsbad, CA, USA). The RNA labeling kit and nucleic acid detection kit were purchased from Roche Diagnostics (Indianapolis, IN, USA).

Suwa H., Kishi H., Imai F., Nakao K., Hirakawa T, & Minegishi T. (2016). Retinoic acid enhances progesterone production via the cAMP/PKA signaling pathway in immature rat granulosa cells. Biochemistry and Biophysics Reports, 8, 62-67.

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Gentamicin sulfate

Lab products found in correlation

78 protocols using gentamicin sulfate

Cell Culture Protocol for CHO-K1, HEK293T, and BHK-21 Cells

Cell Culture Protocol for CHO-K1, HEK293T, and BHK-21 Cells

Characterization of Mouse Tumor Cell Lines

Characterization of Mouse Tumor Cell Lines

Culturing Breast, Oral Squamous Cell Lines

Cell Culture and Palbociclib Sensitivity

Isolation of Lymphocytes from Primate Samples

Cell Culture Conditions for Various Cell Lines

MAYV Virus Propagation in Cell Lines

Cellular Signaling Pathway Analysis

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