Rpmi 1640 media

Manufactured by Sartorius

Sourced in Israel, Switzerland

RPMI-1640 media is a commonly used cell culture medium. It is designed to support the growth and maintenance of a variety of cell types, including mammalian cells, in in vitro settings.

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Lab products found in correlation

Fetal bovine serum (fbs), by Sartorius (5 mentions) Collagenase type 3, by Worthington (2 mentions) Dnase 1, by Roche (2 mentions) Collagenase 4, by Worthington (2 mentions) L glutamine, by Sartorius (2 mentions) Rpmi 1640 media, by Roche (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Penicillin, by Solarbio (1 mentions) Streptomycin, by Solarbio (1 mentions) Dmem media, by Sartorius (1 mentions) Osimertinib, by Selleck Chemicals (1 mentions) Apatinib, by Selleck Chemicals (1 mentions) Ficoll paque plus, by GE Healthcare (1 mentions) Crl 2974, by American Type Culture Collection (1 mentions) Crl 2975, by American Type Culture Collection (1 mentions) Dxp athena flow cytometer, by Cytek (1 mentions) Minimum essential medium (mem), by Sartorius (1 mentions) Rbl 2h3 cells, by American Type Culture Collection (1 mentions) Gentamicin, by Sartorius (1 mentions) Penicillin streptomycin, by Sartorius (1 mentions)

Spelling variants of this product

RPMI-1640 (255 citations) RPMI media (4 citations) RPMI-1640 culture media (2 citations)

15 protocols using rpmi 1640 media

Isolation of PBMCs and lymphocytes

Cited 1 time

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Venous blood was collected from all participants. PBMCs were isolated from the whole blood using Human PBMC Separation Medium (TBD Science, China), centrifuged (Eppendorf, Germany) following the manufacturer’s instruction (TBD Science, China), and resuspended in PBS with 0.5% BSA.
For the isolation of lymphocytes from the lung and colon, the single-cell suspension was obtained according to a previous study.39 (link) Fresh lung tissues were minced and incubated with 1 mg/mL collagenase IV (Worthington, USA) and 50 μg/mL DNase I (Roche, Switzerland) in RPMI-1640 media (Biological Industries, Israel) before being mashed through 70 μm cell strainers. After removing adherent fat tissue and Peyer’s patches, the colon was washed twice with 20 mL HBSS medium containing 5 mM EDTA and 1 mM DTT to remove epithelial cells. Next, 2 mg/mL collagenase type III (Worthington, USA) and 50 μg/mL DNase I (Roche, Switzerland) in RPMI-1640 media were used to digest colon tissues. The digested tissues were filtered through 70 μm cell strainers to obtain cell suspension and enriched with a 40% Percoll gradient after red blood cells were lysed. PBMCs and single-cell suspensions from all tissues were used for subsequent flow cytometry staining.

Wang Y., Li N., Li Q., Liu Z., Li Y., Kong J., Dong R., Ge D., Li J, & Peng G. (2021). Xuanbai Chengqi Decoction Ameliorates Pulmonary Inflammation via Reshaping Gut Microbiota and Rectifying Th17/Treg Imbalance in a Murine Model of Chronic Obstructive Pulmonary Disease. International Journal of Chronic Obstructive Pulmonary Disease, 16, 3317-3335.

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HL-60 and K562 Cell Culture

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HL-60 (1101HUM-PUMC000037) and K562 (1101HUM-PUMC000039) cells were sourced from the National Infrastructure of Cell Line Resource. No mycoplasma contamination was detected in the above cell lines. The cells were maintained in RPMI-1640 media (Biological Industries, Kibbutz Beit-Haemek, Israel) supplemented with 10% fetal bovine serum (FBS, Gibco), 100 U/mL penicillin, and 10 mg/mL streptomycin (Solarbio, Beijing, China) at 37 °C with 5% CO₂. HL-60 cells transduced with luciferase were used in some experiments as indicated.

Wu X., Jiao Z., Zhang J., Li F, & Li Y. (2023). Expression of TFRC helps to improve the antineoplastic effect of Ara-C on AML cells through a targeted delivery carrier. Journal of Nanobiotechnology, 21, 126.

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NSCLC Cell Lines and Targeted Therapies

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The human NSCLC cell lines PC9 (exon 19 deletion) and H1975 (exon 21 L858R and exon 20 T790M) were obtained from the American Type Culture Collection. Cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) media (Biological Industries, Kibbutz Beit Haemek, Israel) or RPMI 1640 media (Biological Industries), supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin at 37 °C in 5% carbon dioxide. Osimertinib and apatinib were purchased from Selleck Chemicals (Shanghai, China).

Liu Y., Xiong Z.C., Sun X., Sun L., Zhang S.L., Ma J.T, & Han C.B. (2019). Impact of apatinib in combination with osimertinib on EGFR T790M-positive lung adenocarcinoma. Translational Cancer Research, 8(5), 2151-2163.

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Isolation of Human Eosinophils from Peripheral Blood

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Peripheral blood samples for eosinophil isolation were collected into sterile vacutainers with ethylenediaminetetraacetic acid (EDTA). Polymorphonuclear leukocytes (PMNs) were isolated by high density gradient centrifugation. The whole blood was layered on Ficoll-Paque PLUS (GE Healthcare, Finland) and centrifuged at 1000 g for 30 min at room temperature. PMNs were separated by hypotonic lysis of erythrocytes and eosinophils were separated using a magnetic eosinophil isolation kit (Miltenyi Biotek, USA). Isolated eosinophils were diluted in cell culture RPMI 1640 media (Biological Industries, Israel) at a final concentration of 2 × 10⁶/mL. The viability of eosinophil was checked flow cytometrically using propidium iodide (2 mg/mL) and it always was > 95 %.

Lavinskiene S., Malakauskas K., Jeroch J., Hoppenot D, & Sakalauskas R. (2015). Functional activity of peripheral blood eosinophils in allergen-induced late-phase airway inflammation in asthma patients. Journal of Inflammation (London, England), 12, 25.

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Culturing Multiple Myeloma Cell Lines

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Human MM cell lines ARP1 and H929 were kind gifts from Prof. Zhiqiang Liu (Department of Physiology and Pathophysiology, School of Basic Medical Science, Tianjin Medical University). MM.1S and MM.1R cells were purchased from ATCC (CRL-2974 and CRL-2975, respectively). Mouse 5TMM3VT cells were donated by Dr. Wen Zhou (Xiangya School of Medicine, Central South University, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education; Key Laboratory of Carcinogenesis, National Health and Family Planning Commission, Changsha, China). The cells were maintained at 37 °C with 5% CO₂ in RPMI 1640 media (#05-065-1A, Biological Industries, Beit Haemek, Israel) supplemented with 10% heat-inactivated fetal bovine serum (FBS; #04-002-1A, Biological Industries, Israel) and 1% penicillin/streptomycin.

Li R., Ke M., Qi M., Han Z., Cao Y., Deng Z., Qian J., Yang Y, & Gu C. (2022). G6PD promotes cell proliferation and dexamethasone resistance in multiple myeloma via increasing anti-oxidant production and activating Wnt/β-catenin pathway. Experimental Hematology & Oncology, 11, 77.

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HepG2 Cell Culture Protocol

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HepG2 cells (ATCC HB-8065) or immortalized mouse primary hepatocytes described in [24 (link)] were cultured in RPMI-1640 media (Biological Industries, Beit HaEmek, Israel) containing 10% Fetal Bovine Serum (FBS; Cat# 12657, Gibco Biosciences, Dublin, Ireland) at 37 °C in a humidified atmosphere of 5% CO₂/95% air. Cell experiments were conducted at > 80% confluence in 6-well plates (7.5 × 10⁵ cells/well) as follows:

Azar S., Udi S., Drori A., Hadar R., Nemirovski A., Vemuri K.V., Miller M., Sherill-Rofe D., Arad Y., Gur-Wahnon D., Li X., Makriyannis A., Ben-Zvi D., Tabach Y., Ben-Dov I.Z, & Tam J. (2020). Reversal of diet-induced hepatic steatosis by peripheral CB1 receptor blockade in mice is p53/miRNA-22/SIRT1/PPARα dependent. Molecular Metabolism, 42, 101087.

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Isolation and Culture of Mast Cells from Mouse Bone Marrow

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RBL-2H3 cells (ATCC, Manassas, VA) were cultured in MEM (Biological Industries, Israel) with 15% foetal bovine serum, 100 U/mL penicillin, and 100 mg/mL streptomycin at 37°C with 5% CO2 in a humidified atmosphere.
Bone marrow mononuclear cells (BMMCs) were derived from female C57 mice at approximately 4 to 6 weeks of age. Isolated bone marrow progenitor cells were cultured in RPMI 1640 media (Biological Industries, Israel) supplemented with foetal bovine serum (FBS, 10%), penicillin (100 U/mL), streptomycin (100 mg/mL), L-glutamine (2 mM), sodium pyruvate (1 mM), HEPES (10 mM) and recombinant cytokines (stem cell factor, 20 ng/mL; IL-3, 20 ng/mL) at 37°C with 5% CO2 in a humidified atmosphere. After 4 weeks, cultures were composed mainly of MCs (90.1%), as the survival rate and differentiation rate of BMMCs were determined by the Cytek Dxp Athena flow cytometer. Analysis of acquired data was performed with FlowJo software. Analysis of the stained populations was performed by gating on the single (FSCW versus FSC), live cells (FSC versus DYE). Then identify a specific mast cell population expressing both CD117 and the FcϵR (CD117 versus FC) (Figure S1).

Li J., Li L., Liu R., Zhu L., Zhou B., Xiao Y., Hou G., Lin L., Chen X, & Peng C. (2022). Integrative lipidomic features identify plasma lipid signatures in chronic urticaria. Frontiers in Immunology, 13, 933312.

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Metastatic Rat Mammary Adenocarcinoma Protocol

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MADB106 is a selected variant cell line obtained from a pulmonary metastasis of a chemically induced mammary adenocarcinoma (MADB100) in the F344 rat [32 (link)]. MADB106 tumor cells metastasize only to the lungs, a process that is dependent upon NK cells [32 (link)]. The lung tumor retention (LTR) of MADB106 cells is highly indicative of the number of metastases that would have developed weeks later [16 (link), 32 (link)-34 (link)]. Additionally, because the metastatic process of MADB106 is sensitive to NK activity predominantly in the first 24 hours following inoculation [32 (link), 33 ], LTR is more reflective of in vivo NK activity levels than the number of actual metastases [16 (link)]. The MADB106 cell line was maintained in monolayer cultures in complete media (CM) (RPMI-1640 media supplemented with 10% heat-inactivated fetal calf serum (FCS), 50μg/mL of gentamicin, 2mM of L-glutamine, 0.1mM of non-essential amino-acids, and 1mM of sodium pyruvate, Biological Industries, Kibbutz Biet Haemek, Israel) in 100% humidity, 5% CO₂ at 37°C. Cells were removed from the culture flask with trypsin solution (0.25% in PBS), and were washed with CM. This cell line was used for both in vivo assessment of lung tumor retention and ex-vivo examination of NK cytotoxicity.

Levi B., Matzner P., Goldfarb Y., Sorski L., Shaashua L., Melamed R., Rosenne E., Page G.G, & Ben-Eliyahu S. (2016). Stress impairs the efficacy of immune stimulation by CpG-C: Potential neuroendocrine mediating mechanisms and significance to tumor metastasis and the perioperative period. Brain, behavior, and immunity, 56, 209-220.

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Isolation of Myeloid Cells from Lung and Colon

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For the isolation of myeloid cells from the lung and colon, the single-cell suspension was obtained according to a previous study.35 (link) Fresh lung tissues were minced and incubated with 1 mg/mL collagenase IV (Worthington, USA) and 50 μg/mL DNase I (Roche, Switzerland) in RPMI-1640 media (Biological Industries, Israel) before being mashed through 70 μm cell strainers. After removing adherent fat tissue and Peyer’s patches, the colon was washed twice with 20 mL HBSS medium containing 5 mM EDTA and 1 mM DTT to remove epithelial cells. Next, 2 mg/mL collagenase type III (Worthington, USA) and 50 μg/mL DNase I (Roche, Switzerland) in RPMI-1640 media were used to digest colon tissues. The digested tissues were filtered through 70 μm cell strainers to obtain cell suspension and enriched with a 40% Percoll gradient after red blood cells were lysed. Single-cell suspensions from all tissues were used for subsequent flow cytometry staining.

Li Y., Li N., Liu J., Wang T., Dong R., Ge D, & Peng G. (2022). Gegen Qinlian Decoction Alleviates Experimental Colitis and Concurrent Lung Inflammation by Inhibiting the Recruitment of Inflammatory Myeloid Cells and Restoring Microbial Balance. Journal of Inflammation Research, 15, 1273-1291.

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Cisplatin-Resistant NSCLC Cell Culture

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Human NSCLC A549 and cisplatin-resistant A549^cisR cells were purchased from the Bogu Biotechnology Company (Shanghai, China) and cultured in RPMI-1640 media (Biological Industries, Israel) supplemented with 10% FBS (Biological Industries) and 1% penicillin/streptomycin (Biological Industries) in a humidified incubator containing 5% CO₂ at 37 °C. For A549^cisR cells, an additional 800 ng/mL cisplatin was added to the media to maintain resistance.
cisplatin and linoleic anhydride were purchased from Tokyo Chemical Industry Co. (Shanghai, China). 7-Ethyl-10-hydroxycamptothecin (SN38) was purchased from Knowshine Pharmachemicals Inc. (Shanghai, China). All other compounds and solvents were purchased from J&K Chemical (Shanghai, China).

Li T., Shi W., Yao J., Hu J., Sun Q., Meng J., Wan J., Song H, & Wang H. (2022). Combinatorial nanococktails via self-assembling lipid prodrugs for synergistically overcoming drug resistance and effective cancer therapy. Biomaterials Research, 26, 3.

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