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Rpmi 1640

Manufactured by Cell Signaling Technology
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RPMI 1640 is a cell culture medium commonly used for the in vitro cultivation of a variety of cell types, including lymphocytes and other hematopoietic cells. It is a complex, serum-supplemented medium that provides essential nutrients, growth factors, and other components required for cell growth and maintenance.

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Lab products found in correlation

Rpmi 1640, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Cell Signaling Technology (1 mentions) Annexin 5 fluorescein isothiocyanate fitc apoptosis detection kit, by BD (1 mentions) Antibodies against bcl 2, by Santa Cruz Biotechnology (1 mentions) Purified colchicine, by Merck Group (1 mentions) Dulbecco s modified eagle s medium dmem, by Cell Signaling Technology (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Propidium iodide, by BD (1 mentions) Novocyte flow cytometer, by Agilent Technologies (1 mentions) Hyclone fetal bovine serum fbs, by GE Healthcare (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Penicillin streptomycin solution, by Corning (1 mentions) Hepg2, by Cell Signaling Technology (1 mentions) Hepg2, by American Type Culture Collection (1 mentions)

5 protocols using rpmi 1640

1

Colchicine-Induced Apoptosis Pathway

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Purified Colchicine (99.89%), MTT, and DMSO were purchased from Sigma–Aldrich Co. (St. Louis, MO, U.S.A.). Antibodies against Bcl-2, Bax, cleaved-caspase-3, p53, Rb, phospho Rb (pRb), β-actin and horseradish peroxidase (HRP) secondary conjugated antibodies were obtained from Santa Cruz Biotechnology, Inc., (Santa Cruz, CA, U.S.A.). Dulbecco’s Modified Eagle’s Medium (DMEM), RPMI 1640, fetal bovine serum (FBS), penicillin, and streptomycin were obtained from Cell Signaling Technology (Danvers, MA, U.S.A.). Annexin V-fluorescein isothiocyanate (FITC) apoptosis detection kit and propidium iodide (PI) were purchased from BD Bioscience (San Jose, CA, U.S.A.).
Yan L., Huang H., Zhang Y., Yuan X., Yan Z., Cao C, & Luo X. (2020). Involvement of p53-dependent apoptosis signal in antitumor effect of Colchicine on human papilloma virus (HPV)-positive human cervical cancer cells. Bioscience Reports, 40(3), BSR20194065.
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2

Isolation and Polarization of CD4+ T-cells

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Normal CD4+ T-cells were isolated from peripheral blood mononuclear cells (PBMCs) using the EasySep (human) negative selection CD4+ T-cell Isolation Kit (Stem cell Technologies, Inc #17951). The CD4+ T-cell purity was analyzed by flow cytometry on an ACEA NovoCyte™ flow cytometer. CD4+ T-cells were maintained in culture at 1x106 cells/mL in RPMI 1640 containing glutamine, 20% FBS, 1% penicillin streptomycin, 1x 2-mercaptoethanol, recombinant human IL-2 (Cell signaling Technology; # 78145; 20 ng/mL). The cells were polarized in IL21 (Cell signaling Technology; #45095; 20ng/ml), and the polarization of CD4+T-cells to TH1 and TH2 was carried out using ImmunoCult™ human TH1 or TH2 (Cell signaling Technology; TH1 (#10975), TH2 (#10973)) differentiation supplement combined with T-cell expansion medium (Cell signaling Technology; #10981), as per the manufacturer’s recommendation. The cells were analyzed on days 7 and 14 for TH1 and TH2 expression patterns, respectively.
Lone W., Bouska A., Sharma S., Amador C., Saumyaranjan M., Herek T.A., Heavican T.B., Yu J., Lim S.T., Ong C.K., Slack G.W., Savage K.J., Rosenwald A., Ott G., Cook J.R., Feldman A.L., Rimsza L.M., McKeithan T.W., Greiner T.C., Weisenburger D.D., Melle F., Motta G., Pileri S., Vose J.M., Chan W.C, & Iqbal J. (2021). Genome-wide microRNA expression profiling of molecular subgroups of peripheral T-cell lymphoma. Clinical cancer research : an official journal of the American Association for Cancer Research, 27(21), 6039-6053.
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3

Mouse B Cell Activation via CD40L Coculture

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The mouse B cells were activated via coculture with mouse fibroblast cells that express membrane-bound CD154/CD40 ligand (CD40L cells) which were a generous gift from Dr. David Sherr (Boston University). CD40L cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% HyClone Fetal Bovine Serum (FBS; GE Healthcare), 100 U/ml of penicillin, 100 μg/mL of streptomycin, 50 μM of 2-mercaptoethanol, and Gibco HT supplement (Thermo Fisher Scientific, Waltham, MA) to select for CD40L-expressing cells. Prior to coculture, the CD40L cells were irradiated (3500 c/Gy) with an X-RAD 320 (PXI, North Branford, CT). Following irradiation, CD40L cells were plated on 96-well plates at a concentration of 5 × 104 cells/mL for ≥24 h. CD40L cells were then washed with RPMI 1640 and isolated B cells were layered on top of the irradiated CD40L cells and supplemented with recombinant interleukin 2 (5 ng/mL, Cell Signaling Technologies, Danver, MA), interleukin 6 (0.0375 ng/mL, Biolegend, San Diego, CA), and interleukin 10 (2 ng/mL, Cell Signaling Technologies). B cells were cocultured with the CD40L cells for 3 days and then an additional 3 days in the absence of the CD40L cells at 37°C and 5% CO2.
Dornbos P., Warren M., Crawford R.B., Kaminski N.E., Threadgill D.W, & LaPres J.J. (2018). Characterizing Serpinb2 as a Modulator of TCDD-Induced Suppression of the B Cell. Chemical research in toxicology, 31(11), 1248-1259.
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4

AMPK Inhibition Impacts Acetyl-CoA Carboxylase

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K562 cells were plated in sterile 6-well plates at 750,000 cells/mL in 4 mL glucose free RPMI-1640 (Gibco) supplemented with 10% FBS (Sigma Aldrich) and 1X penicillin/streptomycin solution (Corning)(3.0 × 106 cells/well) and incubated for 16 h 2-Deoxy-d-glucose was added to a concentration of 12 mM and cells were incubated for 1 h, after which AMPK inhibitors diluted in glucose free RPMI-1640 were added at relevant concentrations and cells were incubated for a further 3 h. Cells were collected, washed with PBS and lysed on ice in lysis buffer containing 1% Triton X (Cell Signaling). Lysates were sonicated (1 × 5 s) and centrifuged for 10 min at 10,000 rpm and 4 °C, and the protein concentration in supernatants was determined by Lowry assay. Lysates were diluted with ELISA PathScan® sample diluent to a final volume of 100 μL and protein concentration of 1.0 mg/ml prior to use. The concentration of p-ACC (Ser79) was determined using an enzyme-linked immunosorbent assay according to recommended protocol (Cell Signaling, ELISA PathScan® phospho-Acetyl-CoA Carboxylase (Ser79)) and expressed as a relative percentage of the DMSO control.
Matheson C.J., Casalvieri K.A., Backos D.S., Minhajuddin M., Jordan C.T, & Reigan P. (2020). Substituted oxindol-3-ylidenes as AMP-activated protein kinase (AMPK) inhibitors. European journal of medicinal chemistry, 197, 112316.
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5

Evaluating Anti-Cancer Drugs on HepG2 Cells

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Cell culture. The human liver cancer cell line HepG2 was purchased from the American Type Culture Collection and maintained in RPMI-1640 (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (FBS; Zhejiang Tian Hang Biological Science and Technology Co., Ltd.), 2.0 g/l sodium hydrogen carbonate, 100 µg/ml streptomycin and 100 U/ml penicillin. The cells were maintained in an incubator at 37˚C (5% CO 2 ), and experiments were conducted during the exponential growth phase. However, the cell line was not authenticated in our laboratory since it was purchased in 2013.
Cell viability assay. HepG2 cells were cultured in RPMI-1640 supplemented with 10% FBS for 24 h. The cells were then treated with TA (Cell Signaling Technology, Inc.) at 0, 90, 180, 270, 360, 450 and 540 µM, and CDDP (Qilu Pharmaceutical Co., Ltd.) at 0, 0.6, 1.2, 1.8, 2.4, 3.0 and 3.6 µg/ml for 24 h. Subsequently, the cells were subjected to MTT analysis according to the manufacturer's protocol (Solar Biotechnology Co., Ltd.). The cell inhibition rate was calculated as follows:
1-(drug A value-control A value)/inhibition rate (%)=1-(drug A value-control A value)/(control A value-blank A value).
Geng N., Zheng X., Wu M., Yang L., Li X, & Chen J. (2019). Tannic acid synergistically enhances the anticancer efficacy of cisplatin on liver cancer cells through mitochondria‑mediated apoptosis. Oncology reports, 42(5).
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