Rpmi 1640

Manufactured by Quality Biological

Sourced in United States

RPMI-1640 is a cell culture medium commonly used for the growth and maintenance of various types of cells, including human and animal cells. It provides the necessary nutrients, vitamins, and salts to support the growth and survival of cells in vitro. RPMI-1640 is a widely used and well-established medium in the field of cell biology and biomedical research.

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Lab products found in correlation

8 protocols using rpmi 1640

Expansion of HIV/EBV-specific CD8+ T cells

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Thawed PBMCs from HIV-infected individuals were labeled with CellTrace Violet (Thermo Fisher Scientific) at 1μM according to the manufacturer's protocol. The cells were then plated at 5 × 10⁶ cells/well in 24 well plate in CellGenix GMP DC Medium (CellGenix) containing recombinant human (rh) IL-7 (1ng/ml, PeproTech) to support minimal T cell survival and rh FLT3L (50ng/ml, PeproTech) to mobilize primary DCs. After 24 hours (day 1), cognate CD8 epitope peptides from HIV or EBV were added to the culture at 1μg/ml. On day 2, human serum (Access Biologicals) and rh IL-2 (Miltenyi Biotec) were added at final concentrations of 8% (volume/volume) and 20U/ml, respectively. Half of the medium was replaced with RPMI-1640 containing 8% human serum, rhIL-2 (20U/ml), and penicillin-streptomycin (100 U/ml, Quality Biological) on days 5, 7 and 9. HIV and EBV-specific CD8⁺ T cells in the culture were analyzed by flow cytometry on days 6 and 12.

Takata H., Kakazu J.C., Mitchell J.L., Kroon E., Colby D.J., Sacdalan C., Bai H., Ehrenberg P.K., Geretz A., Buranapraditkun S., Pinyakorn S., Intasan J., Tipsuk S., Suttichom D., Prueksakaew P., Chalermchai T., Chomchey N., Phanuphak N., de Souza M., Michael N.L., Robb M.L., Haddad E.K., Crowell T.A., Vasan S., Valcour V.G., Douek D.C., Thomas R., Rolland M., Chomont N., Ananworanich J, & Trautmann L. (2022). Long-term antiretroviral therapy initiated in acute HIV infection prevents residual dysfunction of HIV-specific CD8+ T cells. eBioMedicine, 84, 104253.

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Culturing Rat Pheochromocytoma PC-12 Cells

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The PC-12 pheochromocytoma cell line derived from rat adrenal medulla was obtained from American Type Culture Collection (Manassas, VA). RPMI-1640, trypsin solution, phosphate-buffered saline, fetal bovine serum (FBS), sodium pyruvate (0.1 M), L-glutamine (0.2 M) and penicillin/streptomycin solution (containing 10,000 units·ml⁻¹ penicillin and 10,000 μg·ml⁻¹ streptomycin) were obtained from Quality Biological (Gaithersburg, MD), horse serum (heat inactivated) was purchased from Biosource (Rockville, MD) and HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) buffer [1 M, pH 7.4] was obtained from Mediatech, Inc. (Manassas, VA). The PC-12 cells were maintained in RPMI-1640 supplemented with 1 mM HEPES buffer, 10% horse serum, 5% FBS, 1% sodium pyruvate, 1% L-glutamine and 1% penicillin/streptomycin.

Paul R.K., Singh N.S., Khadeer M., Moaddel R., Sanghvi M., Green C.E., O’Loughlin K., Torjman M.C., Bernier M, & Wainer I.W. (2014). (R,S)-Ketamine metabolites (R,S)-norketamine and (2S,6S)-hydroxynorketamine increase the mammalian target of rapamycin (mTOR) function. Anesthesiology, 121(1), 149-159.

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Cell culture conditions for coronavirus

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The cell lines used in this study were purchased from ATCC: MRC-5 (human lung epithelial cell; ATCC: CCL-171), HCT-8 (human ileocecal adenocarcinoma cell; ATCC: CCL-244), Mv1Lu (mink lung epithelial cell; ATCC: CCL-64), RD (human rhabdomyosarcoma cell; ATCC: CCL-136), LLC-MK2 (rhesus monkey kidney cell; ATCC: CCL-7), Vero (African green monkey kidney cell; ATCC: CCL-81). HCoV-229E (Catalog No. FR-303) and OC43 (Catalog No. FR-302) were obtained from International Reagent Resources (IRR) followed by propagation in MRC-5 and HCT-8 cells for 3–5 days post infection, respectively.
Medium for MRC-5, Mv1Lu, RD, LLC-MK2, and Vero cells was DMEM (Cat# 112-300; Quality Biological, Gaithersburg, MD, USA) supplemented with 10% FBS, 1× MEM nonessential amino acids (NEAA; Cat# 25-025; Corning, Corning, NY, USA), 1 mM of sodium pyruvate (Cat# 25-000; Corning, Corning, NY, USA), 2 mM of L-glutamine (Cat# 25-005; Corning, Corning, NY, USA), and 1× penicillin and streptomycin (Cat#30-002; Corning, Corning, NY, USA) except HCT-8 which was cultured with RPMI 1640 (Cat# 112-025; Quality Biological, Gaithersburg, MD, USA) containing 10% FBS.

Bracci N., Pan H.C., Lehman C., Kehn-Hall K, & Lin S.C. (2020). Improved plaque assay for human coronaviruses 229E and OC43. PeerJ, 8, e10639.

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HTLV-1-infected T-cell Line Lysis and Protein Quantification

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C81 is an HTLV-1-infected T-cell line that expresses Tax protein established from patients with T-cell leukemia. These cells are available through AIDS reagent catalog [53] (link)–[55] (link). Cells were cultured in RPMI-1640 containing 10% fetal bovine serum, 1% penicillin/streptomycin, and 1% L-glutamine (Quality Biological) and were incubated in a 5% CO₂ incubator at 37°C. Cells were cultured to confluency and pelleted at 4°C for 15 min at 3,000 rpm. The cell pellets were washed twice with 25 ml of phosphate-buffered saline (PBS) without Ca²⁺ and Mg²⁺ (Quality Biological) and centrifuged once more. Cell pellets were resuspended in lysis buffer (50 mM Tris–HCl, pH 7.5, 120 mM NaCl, 5 mM EDTA, 0.5% NP-40, 50 mM NaF, 0.2 mM Na₃VO₄, 1 mM DTT, one complete protease cocktail tablet/50 ml) and incubated on ice for 20 min, with a gentle vortexing every 5 min. Cell lysates were transferred to Eppendorf tubes and were centrifuged at 10,000 rpm for 10 min. Supernatants were transferred to a fresh tube where protein concentrations were determined using Bio-Rad protein assay (Bio- Rad, Hercules, CA).

Chen H., Zhao Y., Li H., Zhang D., Huang Y., Shen Q., Van Duyne R., Kashanchi F., Zeng C, & Liu S. (2014). Break CDK2/Cyclin E1 Interface Allosterically with Small Peptides. PLoS ONE, 9(10), e109154.

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CHO Cell Culture Protocol

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Chinese hamster ovary (CHO) E77.4 cells (34 (link)) purchased from ATCC were cultured in media supplemented with RPMI 1640 (Quality Biological 112-025-101), 10% heat inactivated fetal bovine serum (FBS) (Gibco 10082147), 2mM L-Glutamine (Quality Biological 118-084-721) and 100 U/mL Penicillin with 100 µg/mL Streptomycin (Quality Biological 120-095-721). Cells were passaged using 0.05% Trypsin-0.1% EDTA (Quality Biological 118-087-721). CHO cells at passages 9-12 were used for all experiments.

Waghela I.N., Mallory K.L., Taylor J.A., Schneider C.G., Savransky T., Janse C.J., Lin P.J., Tam Y.K., Weissman D, & Angov E. (2022). Exploring in vitro expression and immune potency in mice using mRNA encoding the Plasmodium falciparum malaria antigen, CelTOS. Frontiers in Immunology, 13, 1026052.

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Culturing and Maintaining PC-12 Cells

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The PC-12 cell line, which is derived from rat adrenal medulla, was obtained from American Type Culture Collection (Manassas, VA, USA). The PC-12 cells were maintained in RPMI-1640 (Quality Biological, Gaithersburg, MD, USA) supplemented with 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer [1 mM, pH 7.4] (Mediatech, Inc., Manassas, VA, USA), 10% heat-inactivated horse serum (Biosource, Rockville, MD, USA), 5% fetal bovine serum (FBS), 1% sodium pyruvate, 5% L-glutamine and 1% penicillin/streptomycin all purchased from Quality Biological.

Singh N.S., Rutkowska E., Plazinska A., Khadeer M., Moaddel R., Jozwiak K., Bernier M, & Wainer I.W. (2016). Ketamine Metabolites Enantioselectively Decrease Intracellular D-Serine Concentrations in PC-12 Cells. PLoS ONE, 11(4), e0149499.

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Inducible MYCN Expression in Neuroblastoma

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NBL cell line MYCN-3 with a tetracycline-controlled MYCN expression construct was cultured and induced at 9 hours as described by Wei et al [57 (link)]. IMR32 were cultured in EMEM, IMR5 and SK-N-AS were cultured in RPMI-1640 (Quality Biological) supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 units/mL penicillin, and 100 µg/mL streptomycin at 37°C in 5% CO₂. Cell lines were routinely checked for mycoplasma using the MycoAlert Mycoplasma Detection Kit (Lonza, Walkersville, MD).

Zhang S., Wei J.S., Li S.Q., Badgett T.C., Song Y.K., Agarwal S., Coarfa C., Tolman C., Hurd L., Liao H., He J., Wen X., Liu Z., Thiele C.J., Westermann F., Asgharzadeh S., Seeger R.C., Maris J.M., Auvil J.M., Smith M.A., Kolaczyk E.D., Shohet J, & Khan J. (2015). MYCN controls an alternative RNA splicing program in high-risk metastatic neuroblastoma. Cancer letters, 371(2), 214-224.

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Culturing Mammary Carcinoma Cell Lines

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The 4T1‐eGFP‐Puro (Imanis Life Sciences) mammary carcinoma cell line was cultured in RPMI‐1640 media (Quality Biological) supplemented with 10% heat‐inactivated exosome‐free foetal bovine serum (FBS) (Peak Serum), 2 mM L‐glutamine (Quality Biological), 100 ug/ml streptomycin (Quality Biological), 100 U/ml penicillin (Quality Biological), and 2 ug/ml puromycin (Invivogen). The MDA‐MBA‐231 (ATCC) adenocarcinoma cell line was cultured in Leibovitz's L‐15 (ATCC) media supplemented with 10% heat‐inactivated exosome‐free foetal bovine serum (FBS) (Peak Serum), 2 mM L‐glutamine (Quality Biological), 100 ug/ml streptomycin (Quality Biological), and 100 ug/ml penicillin (Quality Biological) for 5 days in a flask (at 37°C and 5% CO2) before harvesting for downstream experiments. For the SLN EV immunogen priming model, the 4T1 (ATCC) mammary carcinoma cell line was cultured in RPMI‐1640 (Quality Biological). When reaching confluency, cells were removed by scraping and resuspended in media after cell viability was determined by Trypan Blue exclusion.

Howard M., Erickson J., Cuba Z., Kim S., Zhou W., Gade P., Carter R., Mitchell K., Branscome H., Siddhi D., Alanazi F., Kim Y., Araujo R.P., Haymond A., Luchini A., Kashanchi F, & Liotta L.A. (2022). A secretory form of Parkin‐independent mitophagy contributes to the repertoire of extracellular vesicles released into the tumour interstitial fluid in vivo. Journal of Extracellular Vesicles, 11(7), e12244.

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