Rmpi 1640

Manufactured by Corning

Sourced in United States

RMPI 1640 is a cell culture medium formulation designed for the growth and maintenance of a variety of mammalian cell types. It provides a balanced salt solution, essential amino acids, vitamins, and other nutrients required for cell proliferation and survival.

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11 protocols using rmpi 1640

Evaluation of PD-L1 Expression in Cell Lines

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The mouse colon carcinoma cell line MC38, the Chinese hamster ovary cell line CHO-K1-hPD-L1, the CHO-K1-mPD-L1 line, and the human T lymphocytic leukemia Jurkat cell line were maintained in RMPI 1640 (Corning, NY, USA). All the cells were cultured in medium supplemented with 10% FBS (Biological Industries, Cromwell, CT, USA) and 100 U/mL penicillin/streptomycin (Solarbio, Beijing, China) in an incubator with 5% CO₂ at 37 °C.
We conducted experiments employing CHO-K1-hPD-L1 and CHO-K1-mPD-L1 cells previously constructed in the laboratory. The flow cytometry results indicated high expression of PD-L1 in both CHO-K1-hPD-L1 and CHO-K1-mPD-L1 cells. Subsequently, we employed flow cytometry to assess the expression of PD-L1 in MC38 cells. The results revealed a high level of PD-L1 expression in MC38 cells. Also, the heightened PD-L1 expression in MC38 cells has been substantiated in multiple studies [32 (link),33 (link),34 (link)]. Notably, our experimental paradigm involved the stimulation of Jurkat cells with PHA and PMA, a methodology documented in the literature, known to elicit PD-1 overexpression within Jurkat cells [35 (link)].

Deng P., Dong X., Wu Z., Hou X., Mao L., Guo J., Zhao W., Peng C., Zhang Z, & Peng L. (2024). Development of Glycosylation-Modified DPPA-1 Compounds as Innovative PD-1/PD-L1 Blockers: Design, Synthesis, and Biological Evaluation. Molecules, 29(8), 1898.

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Maintenance of Human Cell Lines

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Human cell lines were purchased from the Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ, Germany) or the American Type Culture Collection (ATCC, US). UCSD-AML1 [MN1-ETV6] were maintained in RMPI-1640 (Corning®, Corning, NY, USA) supplemented with 20% fetal bovine serum (FBS) (Life Technologies, Carlsbad, CA, USA), 50 U/ML Penicillin–Streptomycin (P/S) (Life Technologies, Carlsbad, CA, USA) and 20 ng/mL human IL-6 and 40 ng/mL human GM-CSF (PreproTech Inc., Rocky Hill, NJ, USA). MV4;11 [KMT2A-AFF1], Molm14 [KMT2A-MLLT3], THP1 [KMT2A-MLLT3], OCI-AML3 [NPM1c], and KASUMI-1 cells [RUNX1-RUNX1T1] were maintained in RMPI-1640 supplemented with 10% FBS and 50 U/ml P/S. 293T cells were maintained in DMEM supplemented with 10% FBS and 50 U/ml P/S.
All cells were cultured in a humidified incubator at 37 °C in 5% CO₂.

Libbrecht C., Xie H.M., Kingsley M.C., Haladyna J.N., Riedel S.S., Alikarami F., Lenard A., McGeehan G.M., Ernst P, & Bernt K.M. (2021). Menin is necessary for long term maintenance of meningioma-1 driven leukemia. Leukemia, 35(5), 1405-1417.

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NSCLC Cell Line Culture and Validation

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The A549, H1299, and PC-9 human NSCLC cell lines were obtained from ATCC. A549 and H1299 cell lines were cultured in RMPI-1640 (Corning, New York, USA) supplemented with 10% FBS (Biological Industries, Kibbutz Beit-Haemek, Israel) in an incubator with 5% CO₂ at 37 ℃. The PC-9 cell line was cultured in DMEM (Corning, New York, USA) supplemented with 10% FBS (Biological Industries, Kibbutz Beit-Haemek, Israel). Monthly PCR tests were conducted to confirm that all cell lines were free of contamination with Mycoplasma. Additionally, all cell lines used in the experiment had been passaged less than ten times.

He T., Wang Y., Lv W., Wang Y., Li X., Zhang Q., Shen H.M, & Hu J. (2024). FBP1 inhibits NSCLC stemness by promoting ubiquitination of Notch1 intracellular domain and accelerating degradation. Cellular and Molecular Life Sciences: CMLS, 81(1), 87.

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Immortalized Podocyte Culture and Treatment

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Immortalized human podocytes were cultured in RMPI 1640 (Corning, Tewksbury, MA, USA) supplemented with 10% fetal bovine serum (FBS) (Corning, Tewksbury, MA, USA), 1% 100 X Penicillin Streptomycin L-Glutamine (PSG) (Corning, Tewksbury, MA, USA), and 1% Insulin-Transferrin-Selenium-Ethanolamine (ITS-X) (Gibco, Gaithersburg, MD, USA) [39 (link)]. Proliferating podocytes were cultured in a humidified atmosphere with 5% CO₂ at 33 °C and the media was changed twice per week and cells were passaged at ~70–80% confluence. Differentiation was induced by placing the cells at 37 °C under the same atmospheric conditions, for 14 days. Differentiated cells were treated with puromycin aminonucleoside (PAN) (Sigma-Aldrich, St. Louis, MO, USA) at 25 ug/mL or with pioglitazone (Alfa Aesar, Tewksbury, MA, USA) at 10 μM. Control cells received treatment of vehicle dimethyl sulfoxide (DMSO) (Sigma-Aldrich, St. Louis, MO, USA). Cells were harvested after 24 h and total RNA isolated.

Bryant C., Webb A., Banks A.S., Chandler D., Govindarajan R, & Agrawal S. (2022). Alternatively Spliced Landscape of PPARγ mRNA in Podocytes Is Distinct from Adipose Tissue. Cells, 11(21), 3455.

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Cell Culture Protocols for CRL-5803 and HEK-293T

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CRL-5803 and CRL-5800 were purchased from American Type Culture Collection (ATCC) and cultured in RMPI1640 (10-040-CV, Corning) supplemented with 8% fetal bovine serum (FBS). HEK-293T cells were all cultured in Dulbecco’s modified Eagle’s medium (DMEM) (10-017-CV, Corning) supplemented with 8% FBS. All the mediums were supplemented with 100 units/ml penicillin and 100 μg/ml streptomycin.

Hong H., Yao S., Zhang Y., Ye Y., Li C., Hu L., Sun Y., Huang H.Y, & Ji H. (2020). In vivo miRNA knockout screening identifies miR-190b as a novel tumor suppressor. PLoS Genetics, 16(11), e1009168.

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Culture and Maintenance of Pancreatic Cancer Cell Lines

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PANC-1 cells were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA) and cultured in Dulbecco’s Modified Eagle Medium (DMEM; Gibco, Gran Island, NY, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS; TCB, Tulare, CA, USA), 100 U/mL penicillin and 10 μg/mL streptomycin. Capan-2 cells were kindly provided by Prof. Yong-Yeon Cho, The Catholic University of Korea (Bucheon-si, Korea) and cultured in RMPI1640 (Corning Inc., Corning, NY, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS; TCB, Tulare, CA, USA), 100 U/mL penicillin and 10 μg/mL streptomycin. All cells were maintained in a humidified atmosphere with 5% CO₂ at 37 °C.

Jeong J.H, & Ryu J.H. (2020). Broussoflavonol B from Broussonetia kazinoki Siebold Exerts Anti-Pancreatic Cancer Activity through Downregulating FoxM1. Molecules, 25(10), 2328.

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Lentiviral Library Transduction of CEM Cells

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CEM cells were cultured in RMPI 1640 (from Corning) with 10% FBS (from Corning). To passage library in T cells, we added 25 mL viruses and 120 μg polybrene to 50 million CEM cells. We achieved 10 ng p24 (10⁸ physical viral particles) for every million CEM cells during infection. We washed cells and completely changed media 6 hours post infection. We supplemented the cells with fresh media 3 days post infection and harvested supernatant 6 days post infection. We centrifuged supernatant at 500 × g for 3 minutes to remove the cells and cell debris. The rest of supernatant was frozen at -80°C.
In summary, we carefully controlled the experiment scales to ensure the library complexity was maintained in every step. Briefly, we harvested >3 × 10⁴E. coli colonies during bacteria transformation, which ensured ∼6-fold coverage of the expected complexity (4608 genotypes). We then transfected 2 × 10⁷ HEK 293T cells with 16 μg plasmid library to package infectious viruses. We used 25 mL viruses (500 ng p24, ∼ 5 × 10⁹ viral particles) to infect 2 × 10⁷ million CEM cells for each biological replicate.

Zhang T.H., Dai L., Barton J.P., Du Y., Tan Y., Pang W., Chakraborty A.K., Lloyd-Smith J.O, & Sun R. (2020). Predominance of positive epistasis among drug resistance-associated mutations in HIV-1 protease. PLoS Genetics, 16(10), e1009009.

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Gastric Cancer Cell Line Cultivation

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Human GC cell lines NCI‐N87, MGC803, HGC27, MKN45, SGC7901, NUGC4, AGS, mouse GC cell line MFC (originally induced from the gastric tissue of 615‐line mice), HEK cell line 293 T, human embryonic lung diploid fibroblasts 2BS, and HUVECs were obtained from Cell Bank of Shanghai Institute of Biochemistry and Cell Biology. All cells were cultured in RMPI‐1640 (Corning) containing 10% FBS and 1% penicillin as well as streptomycin at 37°C under a humidified atmosphere of 5% CO₂.

Zhang Y., Liang K., Zhou X., Zhang X., Xu H., Dai H., Song X., Yang X., Liu B., Shi T, & Wei J. (2023). Combination therapy of DKK1 inhibition and NKG2D chimeric antigen receptor T cells for the treatment of gastric cancer. Cancer Science, 114(7), 2798-2809.

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Culturing Human Prostate Cancer Cell Lines

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22Rv1 (Cat # CRL-2505) and VCaP (Cat # CRL-2876) cells were obtained from ATCC. ATCC authenticates human cancer cell lines using short tandem repeat analysis. CWR-R1-D567 cells were kindly gifted to us from Dr. Scott Dehm (University of Minnesota)^{40 (link)}. These cell lines were expanded, and early passages were frozen in liquid nitrogen. 22Rv1 and CWR-R1-D567 were maintained in RMPI1640 (Corning) supplemented with 10% FBS, 100 U/mL penicillin, and 100 μg/mL streptomycin (penicillin/streptomycin) in a 5% CO₂ incubator at 37 °C. VCaP cells were cultured in DMEM (Corning) with 10% FBS and penicillin/streptomycin in a 5% CO₂ incubator at 37 °C.

Gjyrezi A., Galletti G., Zhang J., Worroll D., Sigouros M., Kim S., Cooley V., Ballman K.V., Ocean A.J., Shah M.A., Scandura J.M., Sboner A., Nanus D.M., Beltran H., Tagawa S, & Giannakakou P. (2021). Androgen receptor variant shows heterogeneous expression in prostate cancer according to differentiation stage. Communications Biology, 4, 785.

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Characterization of BRAF-mutated Melanoma Cell Lines

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BRAF-mutated melanoma cell lines used in this study were obtained from the Massachusetts General Hospital Cancer Center with the following primary sources: COLO858 (ECACC) and MMACSF (Riken Bioresource Center). Each cell line was independently authenticated by Short Tandem Repeats (STR) profiling by ATCC. COLO858 cells were grown in RMPI 1640 (Corning cellgro, Cat. 10–040 CV), and MMACSF cells were grown in DMEM/F-12 (Thermo Fisher Scientific, Cat. 11330–032). For both cell lines, growth media were supplemented with 5% fetal bovine serum (Thermo Fisher Scientific, Cat. 26140–079) and 1% sodium pyruvate (Thermo Fisher Scientific, Cat. 11360–070). We added penicillin and streptomycin at 100 U/ml (Thermo Fisher Scientific, Cat. 15140–122) and plasmocin at 0.5 μg/ml (InvivoGen, Cat. ant-mpp) to all growth media. Cells were engineered to stably express H2B-Venus and mCherry-Geminin fluorescent reporters as described previously [21 (link)]. Engineered and parental cell lines were confirmed to grow at comparable rates in the absence of any treatment or in the presence of different concentrations of BRAF inhibitor Vemurafenib over 72 hours of treatment.

Comandante-Lou N., Khaliq M., Venkat D., Manikkam M, & Fallahi-Sichani M. (2020). Phenotype-based probabilistic analysis of heterogeneous responses to cancer drugs and their combination efficacy. PLoS Computational Biology, 16(2), e1007688.

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