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Penicillin streptomycin l glutamine

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Penicillin/streptomycin/L-glutamine is a common laboratory reagent used as a cell culture supplement. It provides a combination of antibiotics to prevent bacterial contamination and the amino acid L-glutamine to support cell growth and proliferation.

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Lab products found in correlation

Fetal bovine serum (fbs), by Corning (10 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions) Mln4924, by Active Motif (4 mentions) Fetal bovine serum (fbs), by Atlanta Biologicals (3 mentions) Dulbecco modified eagle medium (dmem), by Corning (3 mentions) Mg132, by Peptide Institute (3 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions) Fetal bovine serum (fbs), by Omega Scientific (2 mentions) Skov3, by American Type Culture Collection (2 mentions) Universal mycoplasma detection kit, by American Type Culture Collection (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Doxycycline, by Merck Group (2 mentions) Neocarzinostatin, by Merck Group (2 mentions) Pen strep glutamine, by Thermo Fisher Scientific (2 mentions) Opti mem, by Thermo Fisher Scientific (2 mentions) Sodium pyruvate, by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Gmscf, by Thermo Fisher Scientific (1 mentions) Iscove modified dulbecco media (imdm), by Thermo Fisher Scientific (1 mentions)

26 protocols using penicillin streptomycin l glutamine

1

Cell Culture Conditions for Diverse Cell Lines

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Parental cell lines from KP mice were previously established and described (Dimitrova et al., 2016 (link)). HEK293T (female), A549 (male), H2009 (female), cell lines were propagated in DMEM at 37°celsius. All media were supplemented with 10% fetal bovine serum (FBS) (Corning Life Sciences) and 1% penicillin/streptomycin/L-glutamine (Corning Life Sciences). For A549 or H2009 cells stably infected with pTRIPZ vectors, KP cells stably infected with pTRIPZ and pLVX-mCherry vectors, and A549 or KP or KPK infected with mirE vectors, cells were propagated in DMEM supplemented with 10% Tet system-approved FBS (Takara/Clontech Laboratories) and 1% penicillin/streptomycin/L-glutamine (Corning Life Sciences). Doxycycline (Sigma-Aldrich) was used at 0.1 μg/mL, Cycloheximide (Sigma-Aldrich) at 100 μg/mL, MLN4924 (Active Biochem) at 2 μM, MG132 (Peptides International) at 10 μM, Hemin (Sigma-Aldrich) at 10 μM, Tin protoporphyrin IX dichloride (TinPPIX) (Tocris) at 10 μM, Ki696 (provided by Craig Thomas lab) at 1 μM. DMSO was used as vehicle treatment in both transwell and scratch assay. Cells were periodically screened for Mycoplasma contamination. No cell lines used in this study were found in the database of commonly misidentified cell lines that is maintained by ICLAC and NCBI Biosample. Specific details about cell lines used are provided in the Key Resource Table.
Lignitto L., LeBoeuf S.E., Homer H., Jiang S., Askenazi M., Karakousi T.R., Pass H.I., Bhutkar A.J., Tsirigos A., Ueberheide B., Sayin V.I., Papagiannakopoulos T, & Pagano M. (2019). Nrf2 activation promotes lung cancer metastasis by inhibiting the degradation of Bach1. Cell, 178(2), 316-329.e18.
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2

Cell Culture Protocols for HEK293FT and CHO-K1

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HEK293FT (Invitrogen) cells were maintained in Dulbecco’s modified Eagle medium (DMEM, Corning) supplemented with 10% FBS (Corning), 1% Penicillin–Streptomycin-l-Glutamine (Corning), and 1% MEM Non-Essential Amino Acids (Gibco). CHO-K1 cells (ATCC) were maintained in Ham’s F-12K (Kaighn’s) medium supplemented with 10% FBS (Corning), 1% Penicillin–Streptomycin-l-Glutamine (Corning), and 1% MEM Non-Essential Amino Acids (Gibco). Doxycycline Hyclate (Dox, Sigma-Aldrich) was diluted in water at 10 mg/μL, stored at −20 ∘C, and used at a final concentration of 4 μM for experiments. Trichostatin A (TSA, Sigma-Aldrich) was diluted in DMSO, stored at −20 ∘C, and used at a final concentration of 100 nM for experiments.
DiAndreth B., Wauford N., Hu E., Palacios S, & Weiss R. (2022). PERSIST platform provides programmable RNA regulation using CRISPR endoRNases. Nature Communications, 13, 2582.
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3

Generation of bone marrow-derived DCs

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To generate CD11c+MHCII+ bone marrow-derived DCs (BMDCs), bulk bone marrow cells were plated (5 × 105 cells/well in 24-well plates; 1 × 105 cell/well in 96-well plates; 35 × 106 cells/well in 15 cm plates) and cultured in IMDM (Gibco) supplemented with 10% heat-inactivated FBS (Atlanta Biologicals), penicillin–streptomycin–l-glutamine (Cellgro), and 10 ng/ml GMSCF (Peprotech) at 37°C in 6% CO2. Media was changed on day 3 and 6 and cells were experimented on day 7 or 8. For splenocyte DCs, we used Miltenyi CD11c-magnetic bead isolation kit following the manufacturers protocol (catalog no. 130-052-001; Miltenyi), and isolated cells were plated in 96-well plates for 24 h. For all experiments, cells were pretreated with IF10 media ± 0.5 μM PLX7904 for 60 min. To stimulate cells, half of the media was removed and replaced with IF10 ± LPS media (2× is 200 μg/ml) for various time points.
Minichino D., Lv K., Chu N., Tong W, & Behrens E.M. (2022). BRAF-V600E utilizes posttranscriptional mechanisms to amplify LPS-induced TNFα production in dendritic cells in a mouse model of Langerhans cell histiocytosis. Journal of leukocyte biology, 112(5), 1089-1104.
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4

HEK-293T and NTM-5 Cell Culture

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Human embryonic kidney (HEK-293T; Life Technologies) and normal TM (NTM-5; kind gift from Abbot Clark, University of North Texas Health Science Center) cells were cultured in low-glucose (1 g/L) Dulbecco's modified Eagle's medium (DMEM; Cellgro, Manassas, VA, USA) supplemented with 10% fetal bovine serum (FBS; Omega Scientific, Tarzana, CA, USA) and 1% penicillin/streptomycin/L-glutamine (Cellgro). All cells were incubated at typical growth conditions (37°C at 5% CO2) unless otherwise indicated and passaged routinely to propagate cells and maintain appropriate confluency.
Zadoo S., Nguyen A., Zode G, & Hulleman J.D. (2016). A Novel Luciferase Assay For Sensitively Monitoring Myocilin Variants in Cell Culture. Investigative Ophthalmology & Visual Science, 57(4), 1939-1950.
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5

Myeloid Progenitor Differentiation Assay

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Bone marrow myeloid progenitors were sorted from mice treated with five doses of PBS, CpG1826, IFNγ, or a combination of both CpG1826 and IFNγ, as described above. Myeloid progenitors (500–2000 per well) were cultured in MEM-α media (Gibco) supplemented with 20% heat-inactivated FBS (Atlanta Biologicals) and penicillin–streptomycin–L-glutamine (Cellgro) at 37°C in 6% CO2. GM-CSF, M-CSF, IL-3, and stem cell factor were obtained from Peprotech and used to stimulate myeloid progenitor cell division and differentiation in liquid cultures. All cytokines were used at 5 ng/mL, except GM-CSF was used at 3.3 ng/mL. After seven days in culture, myeloid progenitor cell progeny were harvested and stained for mature myeloid cell surface markers (Ly6G, Ly6C, and CD11b) using the cellular immunophenotyping protocol described above. Total myeloid cell counts were enumerated on a Miltenyi MacsQuant flow cytometer and gated on individual cell populations in FlowJo.
Weaver L.K., Niansheng C, & Behrens E.M. (2018). Toll-like receptor 9 activation potentiates interferon-γ-mediated immunopathology in a murine model of Macrophage Activation Syndrome. Arthritis & rheumatology (Hoboken, N.J.), 71(1), 161-168.
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6

Ovarian Cancer Cell Culture and Carboplatin Treatment

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Fifteen ovarian cancer cell lines were obtained as a gift from the Duke Gynecological Oncology Research Group. These cell lines are shown in Fig. 2. All ovarian cancer cell lines were cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS) (Corning Life Sciences) and 1% penicillin/streptomycin/L-glutamine (Corning Life Sciences).
To generate the omental conditioned media for culture, fresh omenta were isolated from sacrificed C57BL/6 J mice. After 3 washes in PBS to remove residual clots from dissection, omenta were minced and cultured in DMEM without FBS for 48 hours. The adipose tissue was then removed, and the media was passed through a 45 μM filter prior to use in cell culture applications.
Carboplatin (Sigma-Aldrich) was dissolved in water and aliquoted to stock solutions of 15 mM kept at − 20 °C, made fresh prior to cell culture treatment and protected from light. The Carboplatin dosage was 100 μM for all experiments.
Bose S., Yao H., Huang Q., Whitaker R., Kontos C.D., Previs R.A, & Shen X. (2022). Using genetically encoded fluorescent biosensors to interrogate ovarian cancer metabolism. Journal of Ovarian Research, 15, 114.
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7

Targeting DCAF Genes in Cell Lines

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A RNAi ON-TARGETplus SMARTpool Cherry-pick Library was purchased from Dharmacon/Horizon Discovery, including oligonucleotides targeting all DCAF genes17 (see Supplementary Table 4 for details). The library was resuspended in 1× siRNA Buffer (Dharmacon, Horizon Discovery) to a final 2 μM stock concentration. HCT-116 and U-2 OS cells, previously transduced with pBabe-mAzG-CCND1, were reverse transfected in a 96-well plate with siRNA:DharmaFECT 2 (Dharmacon, Horizon Discovery) complex using 20 nM siRNA and 0.1 μl of DharmaFECT 2 per well, according to the manufacturer’s instructions. siRNA:DharmaFECT 2 was complexed in Opti-MEM (Thermo Fisher Scientific). Cells were seeded in FluoroBrite DMEM (Thermo Fisher Scientific) supplemented with 10% FBS (Corning Life Sciences) and 1% penicillin/streptomycin/l-glutamine (Corning Life Sciences). Images were acquired every 6 h for a total of 72 h using a Cytation 5 Imaging Reader (BioTek), equipped with a humidified BioSpa 8 Automated Incubator (BioTek) set at 37 °C and 5% CO2. Phase contrast imaging was used to generate a cell mask. Total fluorescence intensity was calculated at each time point within the cell mask, and normalized to the first time point (time = 0). Images were analysed with Gen5 (version 3.0).
Simoneschi D., Rona G., Zhou N., Jeong Y.T., Jiang S., Milletti G., Arbini A.A., O’Sullivan A., Wang A.A., Nithikasem S., Keegan S., Siu Y., Cianfanelli V., Maiani E., Nazio F., Cecconi F., Boccalatte F., Fenyö D., Jones D.R., Busino L, & Pagano M. (2021). CRL4AMBRA1 is a master regulator of D-type cyclins. Nature, 592(7856), 789-793.
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8

Ovarian Cancer Cell Line Cultivation

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Ovarian cancer cell lines were obtained as described in the key resources table and were cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS) (Corning Life Sciences) and 1% penicillin/streptomycin/L-glutamine (Corning Life Sciences). Cultures were maintained at 37°C in 5% CO2 incubator and were periodically screened for Mycoplasma contamination. HEYA8, SKOV3, and IGROV1 ovarian cancer cells were obtained from ATCC, and no further authentication was performed. The remaining cell lines were obtained and authenticated by the Duke Gynecological Oncology Core Resource, which performed polymorphic short tandem repeat (STR) profiling through the Duke University DNA Analysis Facility.
Bose S., Huang Q., Ma Y., Wang L., Rivera G.O., Ouyang Y., Whitaker R., Gibson R.A., Kontos C.D., Berchuck A., Previs R.A, & Shen X. (2022). G6PD inhibition sensitizes ovarian cancer cells to oxidative stress in the metastatic omental microenvironment. Cell reports, 39(13), 111012.
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9

Cell Culture Conditions Across Cell Lines

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HEK293T (female), 293H (unknown sex), VCAP (male), A375 (female) and SK-MEL-28 (male) cell lines and MEFs (unknown sex) were cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS) (Corning Life Sciences) and 1% penicillin/streptomycin/L-glutamine (Corning Life Sciences). For cells stably infected with doxycycline-inducible vectors, cells were propagated in media supplemented with 10% Tet system-approved FBS (Takara/Clontech Laboratories). RWPE1 (male) cells were cultured with the Keratinocyte Serum Free Medium (K-SFM) supplemented with bovine pituitary extract (BPE) and human recombinant epidermal growth factor (EGF) obtained from Thermo Fisher Scientific (Cat. No. 17005-042). Cells were periodically screened for Mycoplasma contamination. No cell lines used in this study were found in the database of commonly misidentified cell lines that is maintained by ICLAC and NCBI Biosample. Specific details about cell lines used are provided in the Key Resource Table.
Duan S., Moro L., Qu R., Simoneschi D., Cho H., Jiang S., Zhao H., Chang Q., de Stanchina E., Arbini A.A, & Pagano M. (2021). Loss of FBXO31-mediated degradation of DUSP6 dysregulates ERK and PI3K-AKT signaling and promotes prostate tumorigenesis. Cell reports, 37(3), 109870.
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10

Murine Melanoma Cell Lines Protocol

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BrafV600E/+; Pten−/−; and Cdkn2a−/− mouse melanoma cells (YUMMER1.7) were kindly provided by Marcus Bosenberg (Yale). B16-GP33 melanoma cells were kindly provided by Dr. Ananda Goldrath (UCSD). Dartmouth murine mutant malignant melanoma-3A (D4M-3A) were kindly provided by Dr. Francesco Marangoni (UC Irvine). Cell lines were maintained in Corning Dulbecco’s modified eagle’s medium with 10% fetal bovine serum (FBS) (D4M-3A), Corning Iscove’s Modified Dulbecco’s medium with 10% FBS and 1% Corning Penicillin-Streptomycin-L-Glutamine (PSG)(for YUMMER1.7), or Corning Iscove’s Modified Dulbecco’s medium with 10% FBS, 1% (PSG) and 1% geneticin (Gibco) (for B16-GP33). Cell lines were passaged two times per week and underwent a minimum of four passages before injections. To mitigate murine cell line adaptations while in culture, cells were cultured for a maximum of two months in vitro. Early passages were frozen down for future use. No additional cell authentication was performed. All cell lines were free of mycoplasma.
DeRogatis J.M., Viramontes K.M., Neubert E.N., Henriquez M.L., Guerrero-Juarez C.F, & Tinoco R. (2022). Targeting the PSGL-1 Immune Checkpoint Promotes Immunity to PD-1 Resistant Melanoma. Cancer immunology research, 10(5), 612-625.
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