1640 medium

Manufactured by Thermo Fisher Scientific

Sourced in United States, China, United Kingdom, Germany, France, Australia

1640 medium is a sterile, liquid cell culture medium designed for the in vitro cultivation of human cells. It is commonly used in research applications to support the growth and maintenance of various cell types.

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Lab products found in correlation

927 protocols using 1640 medium

Culturing Ovarian Cancer Cell Lines

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The human SKOV3 and A2780 ovarian cancer cell lines and the IOSE80 normal ovarian epithelial cell line were purchased from Wuhan Procell Life Science&Technology (Wuhan, China). The OVCAR-3 cell line was obtained from the Cell Resource Center of Shanghai Institute of Life Sciences Chinese Academy of Sciences (Shanghai, China). The SKOV3 cell line was cultured in McCoy’s 5 A medium (Gibco, Carlsbad, CA, USA), the A2780 cell line was cultured in 1640 medium (Gibco, Carlsbad, CA, USA), and the IOSE80 cell line was cultured in DMEM (Gibco, Carlsbad, CA, USA). The three cell lines were cultured in medium supplemented with 10% fetal bovine serum (Gibco, Carlsbad, CA, USA) and 1% penicillin and streptomycin. The OVCAR-3 cell line was cultured in 1640 medium (Gibco, Carlsbad, CA, USA) supplemented with 20% fetal bovine serum and 0.01 mg/ml insulin. The cells were cultured in an incubator at 37 °C in 5% CO₂.

Zhang X., Gu W., Lin A., Duan R., Lian L., Huang Y., Li T, & Sun Q. (2023). The role of OIP5 in the carcinogenesis and progression of ovarian cancer. Journal of Ovarian Research, 16, 185.

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Culturing Pancreatic Cell Lines

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The pancreatic cancer cell lines BxPC-3, PANC-1, MIA PaCa-2, and AsPC-1 and the immortalized normal pancreatic duct cell line HPDE were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). PANC-1 and MIA PaCa-2 cells were cultured in DMEM (Gibco, USA), and BxPC-3, AsPC-1, HPDE cells were cultured in 1640 medium (Gibco, USA). Both DMEM and 1640 medium were supplemented with 10% FBS (Gibco, USA) and a 1× penicillin-streptomycin solution (Life Technologies, USA). A PSC (Human Pancreatic Stellate Cells, Cat. 3830) cell line was obtained from ScienceCell^TM and maintained in the recommended stellate cell medium. All cells were kept in a humidified incubator at 37°C with 5% CO₂.

Zhang Y.F., Zhou Y.Z., Zhang B., Huang S.F., Li P.P., He X.M., Cao G.D., Kang M.X., Dong X, & Wu Y.L. (2019). Pancreatic cancer-derived exosomes promoted pancreatic stellate cells recruitment by pancreatic cancer. Journal of Cancer, 10(18), 4397-4407.

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Tumor-Infiltrating Lymphocyte Isolation

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Fresh tumor specimens, peripheral blood (n=19) and paraffin-embedded tumor tissues (n=197) were collected from patients newly diagnosed with CC at Sun Yat-Sen University Cancer from 2008 to 2018. The detailed clinical data are described in online supplemental table S1. Peripheral blood was obtained from healthy donors.
TILs were isolated from fresh tumor specimens by mincing, digested by collagenase type IV (Sigma-Aldrich, St. Louis, Missouri, USA) and then cultured in 12-well plates in X-VIVO-15 medium (Lonza, Walkersville, Maryland, USA) for 2–3 days.
Separation of peripheral blood mononuclear cells (PBMCs) was performed by Ficoll (Thermo, Rockford, Illinois, USA) and density gradient centrifugation. The HeLa and SiHa cell lines were maintained in our laboratory in 1640 medium (Gibco, Grand Island, New York, USA) supplemented with 10% fetal bovine serum (Excell Bio, Shanghai, China). The human HEK293T and TC-1 cell lines were purchased from American Type Culture Collection, maintained in our laboratory and cultured in Dulbecco’s Modified Eagle Medium or 1640 medium (Gibco) supplemented with 10% fetal bovine serum. All of the cell lines were subjected to mycoplasma testing and were found to be free of contamination.

Ni H., Zhang H., Li L., Huang H., Guo H., Zhang L., Li C., Xu J.X., Nie C.P., Li K., Zhang X., Xia X, & Li J. (2022). T cell-intrinsic STING signaling promotes regulatory T cell induction and immunosuppression by upregulating FOXP3 transcription in cervical cancer. Journal for Immunotherapy of Cancer, 10(9), e005151.

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Hypoxia Culture of MKN-45 Cells

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MKN-45 cells were kindly provided by Key Laboratory for Experimental Teratology of the Ministry of Education, Department of Pathology, School of Basic Medical Sciences, Shandong University. MKN-45 cells were maintained in 1640 medium (Gibco) supplemented with fetal bovine serum (FBS, 10% PAN), penicillin (100U/mL, Thermo Fisher), and streptomycin (100U/mL, Thermo Fisher) and cultured in 95% air and 5% CO₂ at 37°C. The 1% O₂ stimulation was maintaining MKN-45 cells by 1640 medium (Gibco) supplemented with fetal bovine serum (FBS, 10% PAN), penicillin (100U/mL, Thermo Fisher), and streptomycin (100U/mL, Thermo Fisher) and cultured in 94% N₂, 5% CO_2, and 1% O₂ at 37°C.

Zhu X., Chen H., Li H., Ren H., Ye C., Xu K., Liu J., Du F., Zhang Z., Liu Y., Xie X., Wang M., Ma T., Chong W., Shang L, & Li L. (2023). ITGB1-mediated molecular landscape and cuproptosis phenotype induced the worse prognosis in diffuse gastric cancer. Frontiers in Oncology, 13, 1115510.

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Cell Culture Conditions for U2OS and MCA205 Cell Lines

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U2OS cell lines were cultured in Dulbecco’s modified Eagle’s medium (#41 966-02, Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (#F7524, Sigma Aldrich), 1% non-essential amino acids (#11 140-035, Thermo Fisher Scientific), 1% HEPES (#15630080, Thermo Fisher Scientific) and 1% penicillin/streptomycin (#15140122, Thermo Fisher Scientific). For U2OS HMGB1-GFP, XBP1ΔDBD-venus-RFP-FYVE, GFP-LC3 and GFP-ATF6 0.5 mg/mL G418 (#10 131-27, Thermo Fisher Scientific) was added to the medium. MCA205 cell lines were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (#61870044, Thermo Fisher Scientific) supplemented with identical components. Cells were maintained in a humidified incubator at 37°C with 5% CO₂. Cell culture consumables were purchased from Corning (New York, USA).

Bezu L., Wu Chuang A., Sauvat A., Humeau J., Xie W., Cerrato G., Liu P., Zhao L., Zhang S., Le Naour J., Pol J., van Endert P., Kepp O., Barlesi F, & Kroemer G. (2022). Local anesthetics elicit immune-dependent anticancer effects. Journal for Immunotherapy of Cancer, 10(4), e004151.

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Culturing GC and Colon Cancer Cells

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GC cells such as SGC-7901, MGC-803, BGC-823, MKN-45, and KATO-III and colon cancer cells such as Caco-2 were obtained from The Cell Bank of Type Culture Collection of the Chinese Academy of Sciences (People’s Republic of China). Cells were cultured in Roswell Park Memorial Institute 1640 medium (Thermo Fisher Scientific, Waltham, MA, USA) containing 10% fetal bovine serum (FBS), penicillin (100 U/mL), and streptomycin (100 mg/mL) in an atmosphere of 95% air and 5% CO₂ at 37°C.

Zhang X., Song Y., Song N., Zhang Y., Zhang L., Wang Y., Wang Z., Qu X, & Liu Y. (2017). RANKL/RANK pathway abrogates cetuximab sensitivity in gastric cancer cells via activation of EGFR and c-Src. OncoTargets and therapy, 10, 73-83.

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Activation and Validation of T Cell Markers

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Human peripheral blood mononuclear cells (PBMCs) were obtained from a leukoreduction collar (Brigham and Women’s hospital Crimson Core Laboratory, MA) with gradient centrifugation. Human PBMCs were activated with PHA and cultured in Roswell Park Memorial Institute 1640 medium (ThermoFisher, MA), supplemented with 10% fetal bovine serum, 10 mM HEPES, 0.1 mM non-essential amino acids, 1 mM sodium pyruvate, 100 U/mL penicillin, 100 μg/mL streptomycin, 50 μM 2-ME, and 50 IU/mL rhIL-2 (NCI, MD) for 3 days or 10 days before being used for validating anti-CTLA4 antibody and anti-PD1 antibody production. PHA-activated PMBCs were incubated with purified anti-CTLA4 antibody and/or anti-PD1 antibody at 4 °C for 25 min, then incubated with commercial PE-labeled anti-human CD279 (PD-1) (329920, BioLegend, CA) or PE-labeled anti-human CD152 (CTLA4) (349906, BioLegend, CA). Flow cytometric analysis was done by LSRII Fortessa cytometer (BD Biosciences, CA). Data analysis was done by the FlowJo software (TreeStar Inc, OR) (Fig. 5e).

Cao J., Perez-Pinera P., Lowenhaupt K., Wu M.R., Purcell O., de la Fuente-Nunez C, & Lu T.K. (2018). Versatile and on-demand biologics co-production in yeast. Nature Communications, 9, 77.

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Cell Line Culturing and Maintenance

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NCI-H460, HEK-293T, SPC-A1, HeLa, BEAS-2B, and other cell lines were purchased from the American Type Culture Collection (Manassas, VA). Tumor cell lines were cultured in Roswell Park Memorial Institute 1640 medium (Thermo Fisher Scientific, Rockford, IL). HEK-293T, HeLa, and BEAS-2B cells were cultured in Dulbecco’s modified Eagle’s medium (Thermo Fisher Scientific, Rockford, IL) containing 10% (v/v) fetal bovine serum, GlutaMax, and 100 U/mL penicillin-streptomycin in an incubator (Thermo Fisher Scientific, Rockford, IL) at 37 °C, 5% CO₂. Cells were sub-cultured approximately every 2 days at 80% confluence using 0.25% (w/v) trypsin (Thermo Fisher Scientific, Rockford, IL) at a split ratio of 1:3.

Wang H., Qin M., Liu R., Ding X., Chen I.S, & Jiang Y. (2019). Characterization of A Bifunctional Synthetic RNA Aptamer and A Truncated Form for Ability to Inhibit Growth of Non-Small Cell Lung Cancer. Scientific Reports, 9, 18836.

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Polystyrene-co-maleic anhydride cell culture

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Polystyrene co-maleic anhydride (molecular weight = ~1600), N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDAC), Hank’s balanced salt solution, Roswell Park Memorial Institute (RPMI) 1640 medium, fetal bovine serum (FBS), bovine serum albumin (BSA), and TrypLE express were bought from ThermoFisher Scientific (Dubai, UAE). L-glutamine and an antibiotic solution of penicillin/streptomycin were purchased from (Merck Hertfordshire, UK). All consumable materials such as Petri dishes, conical tubes (15 mL and 50 mL), cell culture flasks (25 cm² and 75 cm²), and dialysis tubing were purchased from (Merck Hertfordshire, 120 Moorgate London, UK).

El-Deeb I.M., Pittala V., Eltayeb D, & Greish K. (2021). Selective Targeting of Breast Cancer by Tafuramycin A Using SMA-Nanoassemblies. Molecules, 26(12), 3532.

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Cell Culture Maintenance and Validation

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All cell lines were cultured in Roswell Park Memorial Institute 1640 Medium (Thermo Fisher Scientific, Cat# 11875-093) with 7% fetal bovine serum (Sigma, Cat# F1051), 1% penicillin/streptomycin (Thermo Fisher Scientific, Cat# 15140-122), and 2 mM L-glutamine (Thermo Fisher Scientific, Cat# 25030-081), except for 293T with Dulbecco’s modified Eagle medium (Thermo Fisher Scientific, Cat# 11995-065) and HEC116 with Eagle’s minimum essential medium (Wisent, Cat# 320-005-CL). Cells were maintained at 37 °C and 5% CO₂ and regularly tested for Mycoplasma using Mycoalert Detection Kit (Lonza, Cat # LT07-318). All cell line origins are listed in Reporting Summary and have been validated by short tandem repeat analysis.

Xue Y., Morris J.L., Yang K., Fu Z., Zhu X., Johnson F., Meehan B., Witkowski L., Yasmeen A., Golenar T., Coatham M., Morin G., Monast A., Pilon V., Fiset P.O., Jung S., Gonzalez A.V., Camilleri-Broet S., Fu L., Postovit L.M., Spicer J., Gotlieb W.H., Guiot M.C., Rak J., Park M., Lockwood W., Foulkes W.D., Prudent J, & Huang S. (2021). SMARCA4/2 loss inhibits chemotherapy-induced apoptosis by restricting IP3R3-mediated Ca2+ flux to mitochondria. Nature Communications, 12, 5404.

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