Penicillin streptomycin l glutamin

Manufactured by Thermo Fisher Scientific

Penicillin/streptomycin/L-Glutamin is a solution that contains the antibiotics penicillin and streptomycin, as well as the amino acid L-glutamine. It is commonly used as a supplement in cell culture media to prevent bacterial contamination and support cell growth.

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6 protocols using penicillin streptomycin l glutamin

Culturing Breast Cancer Cell Lines

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MCF-10A, MCF-7 and SUM-159 were purchased from National Infrastructure of Cell Line Resource of China.
MCF-10A cells were maintained in F12 medium (Gibco) supplemented with 5% horse serum (Gibco), 1%(vol/vol) penicillin/streptomycin/L-Glutamin (Gibco), 10 mg/mL insulin, 20 mg/mL EGF, 100 mg/mL cholera toxin and 0.5 mg/mL hydrocortisone. SUM-159 cells were maintained in F12 medium (Gibco) supplemented with 5% fetal bovine serum (GEMINI), 1% (vol/vol) penicillin/streptomycin/L-Glutamin (Gibco), 5 mg/mL insulin and 10 mg/mL dexamethasone. MCF-7 cells were maintained in Dulbecco’s Modified Eagle Medium (Gibco) supplemented with 10% (vol/vol) fetal bovine serum (GEMINI) and 1% (vol/vol) penicillin/streptomycin/L-Glutamin (Gibco). All cells were cultured at 37°C, 5% CO₂ in a humidified atmosphere.

Tian X., Lu J., Nanding K., Zhang L., Liu Y., Mailisu M., Morigen M, & Fan L. (2022). The antihyperlipidemic drug potassium piperonate impairs the migration and tumorigenesis of breast cancer cells via the upregulation of miR-31. Frontiers in Oncology, 12, 828160.

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Cell Culture of Breast Cancer Lines

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Cell Strains and Cell Culture MCF-10A, MCF-7 and SUM-159 were purchased from National Infrastructure of Cell Line Resource of China.
MCF-10A cells were maintained in F12 medium (Gibco) supplemented with 5% horse serum (Gibco), 1% (vol/vol) penicillin/streptomycin/L-Glutamin (Gibco) ,10 mg/mL insulin, 20 mg/mL EGF, 100 mg/mL Cholera Toxin and 0.5 mg/mL Hydrocortisone. SUM-159 cells were maintained in F12 medium (Gibco) supplemented with 5% fetal bovine serum (GEMINI), 1% (vol/vol) penicillin/streptomycin/L-Glutamin (Gibco), 5 mg/mL insulin and 10 mg/mL dexamethasone. MCF-7 cells were maintained in Dulbecco's Modi ed Eagle Medium (Gibco) supplemented with 10% (vol/vol) fetal bovine serum (GEMINI) and 1% (vol/vol) penicillin/streptomycin/L-Glutamin (Gibco). All cells were cultured at 37ºC, 5% CO 2 in a humidi ed atmosphere.

Lü J., Tian X., Mailisu M., Morigen M., & Fan L. (2021). The antihyperlipidemic drug Potassium Piperate impairs migration and tumorgenesis of breast cancer cells via upregulation of miR-31.

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Cell Culture and Synchronization Protocol

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Gastric cancer cells (MKN45 and BGC-803) were maintained in RPMI 1,640 medium (Gibco) supplemented with 10% fetal bovine serum (BI) and 1% (vol/vol) penicillin/streptomycin/L-Glutamin (Gibco). Human gastric mucosal epithelial cells (GES-1) were maintained in DMEM (High glucose, Gibco) supplemented with 10% (vol/vol) fetal bovine serum (BI) and 1% (vol/vol) penicillin/streptomycin/L-Glutamin (Gibco). All cells were cultured at 37°C and 5% CO₂ in a humidified atmosphere (Thermo Fisher Scientific, MA, United States). To synchronize cells in the G1/S-phase, the cells were cultured in serum-free medium for 24 h.

Zhao H., Gezi G., Tian X., Jia P., Morigen M, & Fan L. (2021). Lysophosphatidic Acid–Induced EGFR Transactivation Promotes Gastric Cancer Cell DNA Replication by Stabilizing Geminin in the S Phase. Frontiers in Pharmacology, 12, 706240.

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Cell Culture and Synchronization

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Cell culture MKN45 and BGC-803 cells were maintained in RPMI 1640 medium (Gibco) supplemented with 10% fetal bovine serum (GEMINI) and 1% (vol/vol) penicillin/streptomycin/L-Glutamin (Gibco), GES-1 cells were maintained in Dulbecco's Modi ed Eagle Medium (Gibco) supplemented with 10% (vol/vol) fetal bovine serum (GEMINI) and 1% (vol/vol) penicillin/streptomycin/L-Glutamin (Gibco). All cells were cultured at 37ºC, 5% CO 2 in a humidi ed atmosphere. To synchronize cells in G1/S-phase, cells were cultured in serum-free medium for 24 hours.

Zhao H., Gezi G., Tian X., Jia P., Morigen M., & Fan L. (2020). Lysophosphatidic Acid-induced EGFR Transactivation Promotes Gastric Cancer DNA Replication Through Up-Regulation of Geminin.

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Transfection of HEK293T Cells with eGFP mRNA

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HEK293T cells were maintained in DMEM medium (Biowest) supplemented with 10% fetal bovine serum (Biowest) and 1% penicillin/streptomycin/L-glutamin (Gibco-Invitrogen) and incubated at 37 °C, 5% CO₂. Twenty-four hours before transfection, cells were seeded in 24-well plates at a density of 150 × 10³ cells per well in 500 μl cell culture medium. Transfections were performed in triplicate and a fixed volume of 16 μl corresponding with a theoretical amount of 0.5 μg eGFP mRNA-loaded LNPs was added to each well to transfect the cells. After 24 h, cells were stained to identify dead cells using a 7-AAD live/dead stain (ThermoFisher) and were analyzed using the MACSQuant 16 flow cytometer (Miltenyi Biotec) to quantify eGFP-expression.

Meulewaeter S., Nuytten G., Cheng M.H., De Smedt S.C., Cullis P.R., De Beer T., Lentacker I, & Verbeke R. (2023). Continuous freeze-drying of messenger RNA lipid nanoparticles enables storage at higher temperatures. Journal of Controlled Release, 357, 149-160.

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Generation of Murine Bone Marrow-Derived Dendritic Cells

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Female C57BL/6 mice (6 weeks old) were purchased from Envigo (Gannat, France) and housed in an SPF facility. All animal experiments were conducted according to the regulations of the Belgian law and approved by the local Ethical Committee. Primary murine bone marrow-derived DC (BM-DC) cultures were generated as previously described. 31 Briefly, bone marrow was isolated from the femurs and tibias and processed into a single cell suspension. After red blood cell lysis, cells were cultured in complete medium (RPMI containing 5% FetalClone TM I (HyClone TM ), 1% penicillin/streptomycin/L-glutamin (Gibco-Invitrogen), β-mercaptoethanol (5 µM, Gibco-Invitrogen) and 40 ng ml -1 GM-CSF (Peprotech) in low-adherence culture dishes. At day 5, all cells were collected by centrifugation, resuspended in the complete medium and seeded for experiments at 5x10 5 cells per well in a 24 well plate.
The mouse melanoma cell line B16-OVA (kindly provided by K. Rock, University of Massachusetts Medical Center) and the T cell lymphoma E.G7-OVA (obtained from the American Type Culture Collection, Rockville, MD, USA) were cultured at 37°C in a humidified 5% CO2 atmosphere in RPMI 1640 medium (Sigma-Aldrich, Diegem, Belgium) supplemented with 10% FBS, 100 U ml -1 penicillin, 100 μg ml -1 streptomycin, 2 mM lglutamine and 0.4 mg ml -1 of the selection agent G418 (Thermo-Scientific, Aalst, Belgium).

Verbeke R., Lentacker I., Breckpot K., Janssens J., Van Calenbergh S., De Smedt S.C, & Dewitte H. (2019). Broadening the Message: A Nanovaccine Co-loaded with Messenger RNA and α-GalCer Induces Antitumor Immunity through Conventional and Natural Killer T Cells. ACS nano, 13(2).

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