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Ramos cells

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Ramos cells are a human lymphoblastoid cell line derived from a Burkitt's lymphoma. They are suspension cells that can be used for various cell biology and biochemistry studies.

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Lab products found in correlation

Pen strep, by Thermo Fisher Scientific (6 mentions) Glutamax, by Thermo Fisher Scientific (5 mentions) Mitotracker orange, by Thermo Fisher Scientific (3 mentions) Rpmi 1640, by American Type Culture Collection (2 mentions) Pen strep, by American Type Culture Collection (2 mentions) Sodium pyruvate, by Thermo Fisher Scientific (2 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (2 mentions) Hek293t cell, by American Type Culture Collection (2 mentions) Rpmi 1640, by Thermo Fisher Scientific (2 mentions) Dulbecco modified eagle medium (dmem), by American Type Culture Collection (2 mentions) Plasmatest, by InvivoGen (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Pe labeled anti mouse igg, by BioLegend (1 mentions) 2 nbdg, by Thermo Fisher Scientific (1 mentions) Cd27 m t271, by BD (1 mentions) Igd ia6 2, by BioLegend (1 mentions) Cd3 hit3a, by BioLegend (1 mentions) 2 nbdg, by American Type Culture Collection (1 mentions) Cd10 hi10a, by BioLegend (1 mentions) Cellrox, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Ramos B cells Ramos cell line B-NHL cell lines (Ramos and CA46

11 protocols using ramos cells

1

Cell Culture Protocols for Diverse Cell Lines

Cited 2 times
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HEK293T cells (ATCC CRL-11268) were cultured in DMEM (ATCC) supplemented with 10% fetal bovine serum (FBS, Atlanta Biologics) and penicillin-streptomycin (Pen/Strep, Gibco).
Jurkat (ATCC TIB-152), J76 cells, and Ramos cells (ATCC CRL-1596) were cultured in RPMI-1640 (ATCC) supplemented with 10% FBS and Pen/Strep. J76 cells88 (link) (PMID 29707134) were a gift from Mirjam Heemskerk and Mark Davis.
CSS-930 TCR library cells were a gift from David Johnson75 (link), and were cultured in RPMI-1640 (ThermoFisher) supplemented with 10% FBS, Pen/Strep, 1x non-essential amino acids (ThermoFisher), 2mM GlutaMAX (ThermoFisher), and 1mM sodium pyruvate (ThermoFisher).
Dobson C.S., Reich A.N., Gaglione S., Smith B.E., Kim E.J., Dong J., Ronsard L., Okonkwo V., Lingwood D., Dougan M., Dougan S.K, & Birnbaum M.E. (2022). Antigen identification and high-throughput interaction mapping by reprogramming viral entry. Nature methods, 19(4), 449-460.
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2

Clearance of Cell Surface Immunocomplexes

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To measure the clearance of immunocomplexes from the cell surface, Ramos cells (ATCC) were incubated with fluorescein isothiocyanate (FITC)-labeled ASKP1240 or FITC-labeled anti-CD40 agonistic mAb (clone G28.5; ATCC) in RPMI1640 supplemented with 10% fetal bovine serum (FBS) for 15 min at 4°C. The stained cells were washed, and then incubated for varying durations in culture medium at 37°C. At the given time points, cells were removed from the culture and washed with phosphate-buffered saline (PBS) containing 0.1% NaN3 to block internalization. Next, the cells were incubated with phycoerythrin (PE)-labeled anti-human IgG (BioLegend, San Diego, CA) or PE-labeled anti-mouse IgG (BioLegend) for 30 min at 4°C. The geometric mean fluorescence intensity (geoMFI) was measured by flow cytometry and analyzed using CellQuest software (ver. 5.1.1; BD Biosciences, Franklin Lakes, NJ).
Okimura K., Maeta K., Kobayashi N., Goto M., Kano N., Ishihara T., Ishikawa T., Tsumura H., Ueno A., Miyao Y., Sakuma S., Kinugasa F., Takahashi N, & Miura T. (2014). Characterization of ASKP1240, a Fully Human Antibody Targeting Human CD40 With Potent Immunosuppressive Effects. American Journal of Transplantation, 14(6), 1290-1299.
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3

Multiparametric Analysis of Immune Cell Function

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Fresh healthy donor human PBMCs were incubated with CellROX (Thermo Fisher Scientific), NAO (Thermo Fisher Scientific), or 2-NBDG (Carbosynth) according to the manufacturer’s specifications. For all three conditions, cells were stained during the efflux incubation for MitoTracker and otherwise prepared for flow cytometry using the protocol detailed above with a few exceptions. 2-NBDG and NAO panels required substitution of MitoTracker Orange (Thermo Fisher Scientific). The fluorochrome-conjugated primary antibody panel was as follows: CD3 (HIT3a, BioLegend), CD19 (SJ25C1, BioLegend), CD27 (M-T271, BD Biosciences), IgD (IA6–2, BioLegend), and CD10 (HI10a, BioLegend). Compensation for the functional dyes required the use of Ramos cells (ATCC), incubated with CellROX, 2-NBDG, or NAO, respectively.
Farmer J.R., Allard-Chamard H., Sun N., Ahmad M., Bertocchi A., Mahajan V.S., Aicher T., Arnold J., Benson M.D., Morningstar J., Barmettler S., Yuen G., Murphy S.J., Walter J.E., Ghebremichael M., Shalek A.K., Batista F., Gerszten R, & Pillai S. (2019). Induction of metabolic quiescence defines the transitional to follicular B cell switch. Science signaling, 12(604), eaaw5573.
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4

EBV Infection of Ramos B Cells

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All cells were confirmed to be free of mycoplasma infection using PlasmaTest (InvivoGen, San Diego, CA). Wild-type EBV was prepared from supernatants of B95–8 cells cultured in RPMI medium 1640 supplemented with 10% FBS for two weeks. Briefly, the cells were pelleted and the virus suspension was filtered through 0.45 μM Millipore filters. The concentrated virus stocks were aliquoted and stored at -80oC.
We infected ~2 x 106 Ramos Cells (ATCC CRL-1596) in the presence of growth medium containing 2μg/ml of phytohemagglutinin (PHA) for 4 hours. The infected cells were washed, cultured in growth media, and observed daily for multinuclear giant cell formation and morphological changes characteristic of EBV-infected B cells. After 10 passages, the infection was confirmed by measuring the expression of viral EBNA2 protein levels (Supplementary Figure 6).
Harley J.B., Chen X., Pujato M., Miller D., Maddox A., Forney C., Magnusen A.F., Lynch A., Chetal K., Yukawa M., Barski A., Salomonis N., Kaufman K.M., Kottyan L.C, & Weirauch M.T. (2018). Transcription factors operate across disease loci, with EBNA2 implicated in autoimmunity. Nature genetics, 50(5), 699-707.
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5

EBV Infection of Ramos B Cells

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All cells were confirmed to be free of mycoplasma infection using PlasmaTest (InvivoGen, San Diego, CA). Wild-type EBV was prepared from supernatants of B95–8 cells cultured in RPMI medium 1640 supplemented with 10% FBS for two weeks. Briefly, the cells were pelleted and the virus suspension was filtered through 0.45 μM Millipore filters. The concentrated virus stocks were aliquoted and stored at -80oC.
We infected ~2 x 106 Ramos Cells (ATCC CRL-1596) in the presence of growth medium containing 2μg/ml of phytohemagglutinin (PHA) for 4 hours. The infected cells were washed, cultured in growth media, and observed daily for multinuclear giant cell formation and morphological changes characteristic of EBV-infected B cells. After 10 passages, the infection was confirmed by measuring the expression of viral EBNA2 protein levels (Supplementary Figure 6).
Harley J.B., Chen X., Pujato M., Miller D., Maddox A., Forney C., Magnusen A.F., Lynch A., Chetal K., Yukawa M., Barski A., Salomonis N., Kaufman K.M., Kottyan L.C, & Weirauch M.T. (2018). Transcription factors operate across disease loci, with EBNA2 implicated in autoimmunity. Nature genetics, 50(5), 699-707.
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6

Cell Culture Protocols for Diverse Cell Lines

Cited 2 times
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HEK293T cells (ATCC CRL-11268) were cultured in DMEM (ATCC) supplemented with 10% fetal bovine serum (FBS, Atlanta Biologics) and penicillin-streptomycin (Pen/Strep, Gibco).
Jurkat (ATCC TIB-152), J76 cells, and Ramos cells (ATCC CRL-1596) were cultured in RPMI-1640 (ATCC) supplemented with 10% FBS and Pen/Strep. J76 cells88 (link) (PMID 29707134) were a gift from Mirjam Heemskerk and Mark Davis.
CSS-930 TCR library cells were a gift from David Johnson75 (link), and were cultured in RPMI-1640 (ThermoFisher) supplemented with 10% FBS, Pen/Strep, 1x non-essential amino acids (ThermoFisher), 2mM GlutaMAX (ThermoFisher), and 1mM sodium pyruvate (ThermoFisher).
Dobson C.S., Reich A.N., Gaglione S., Smith B.E., Kim E.J., Dong J., Ronsard L., Okonkwo V., Lingwood D., Dougan M., Dougan S.K, & Birnbaum M.E. (2022). Antigen identification and high-throughput interaction mapping by reprogramming viral entry. Nature methods, 19(4), 449-460.
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7

Cell Culture Maintenance and Imaging

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Ramos cells were obtained from the American Type Culture Collection. Jurkat cells were a kind gift from A. Weiss (University of California, San Francisco [UCSF]). Both Jurkat and Ramos cells were maintained in 10G-RPMI (RPMI 1640 supplemented with l-glutamine and 25 mM HEPES [Mediatech] and containing 1% [vol/vol] GlutaMAX [Life Technologies] and 10% [vol/vol] fetal bovine serum [FBS; Life Technologies]). Cultures were maintained in a 37°C/5% CO2 incubator at a density ranging from 0.2 to 1 million cells/ml. HEK-293T cells (used to generate lentivirus for transduction) were maintained in DMEM (Mediatech) supplemented with 10% (vol/vol) FBS (Life Technologies). Cells were imaged in imaging media (RPMI 1640 without phenol red supplemented with 25 mM HEPES, 1% l-glutamine, and 0.1% FBS).
Mclaurin J.D, & Weiner O.D. (2019). Multiple sources of signal amplification within the B-cell Ras/MAPK pathway. Molecular Biology of the Cell, 30(13), 1610-1620.
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8

Isolation and Cultivation of Various Immune Cells

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Ramos cells, the human Burkitt’s lymphoma cell line, was purchased from the American Type Culture Collection (Manassas, VA, USA). Primary human B cells were purified from healthy donors using either a RosetteSep Human B Cell Enrichment Cocktail (Stem Cell Technologies, Vancouver, BC, Canada) or a B cell isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany).
Blood was obtained from healthy donors after obtaining informed consent according to the Declaration of Helsinki. This was approved by the Institutional Review Board at Seoul National University Hospital. Human monocytes and plasmacytoid dendritic cells (PDCs) were isolated using human monocyte isolation kit II (Miltenyi Biotec) or human plasmacytoid dendritic isolation kit (Miltenyi Biotec). Murine bone marrow-derived macrophages (BMMs) were generated by culturing murine bone marrow cells in the presence of 10 ng/mL M-CSF (Sigma-Aldrich, St Louis, MO, USA) for 7 days.
Park J.K., Byun J.Y., Park J.A., Kim Y.Y., Lee Y.J., Oh J.I., Jang S.Y., Kim Y.H., Song Y.W., Son J., Suh K.H., Lee Y.M, & Lee E.B. (2016). HM71224, a novel Bruton’s tyrosine kinase inhibitor, suppresses B cell and monocyte activation and ameliorates arthritis in a mouse model: a potential drug for rheumatoid arthritis. Arthritis Research & Therapy, 18, 91.
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9

Ginsenoside Treatment on Lymphoma Cell Lines

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Ramos cells (American Type Culture Collection, Manassas, VA, USA) and the human Burkitt’s lymphoma cell line were cultured in suspension by Roswell Park Memorial Institute (RPMI) 1640 medium (Gibco, Rockville, MD, USA) with 10% fetal bovine serum (Gibco) at 37 °C in 5% CO2 in air. The in vitro-treated ginsenoside compounds (> ~99 %) were obtained from MODNBIO Inc. (Seoul, Korea). The Ibrutinib (PCI-32765) (Selleckchem, Houston, TX, USA) was used as a control.
Lee K.W., Lee W.H., Han B.S., Lee J.H., Doo E.K, & Kim J.H. (2020). Molecular Drug Discovery of Single Ginsenoside Compounds as a Potent Bruton’s Tyrosine Kinase Inhibitor. International Journal of Molecular Sciences, 21(9), 3065.
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10

Complement-Dependent Cytotoxicity Assay for Anti-PD-1 Antibodies

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Complement-dependent cytotoxicity (CDC) refers to the cell lysis effects induced by the membrane attack complex formed by the binding of specific antibodies to the corresponding cell surface antigens, which triggers the canonical complement activation pathway. The activated CD4+ T cells (2×104 cells/well) were used as the target cells, mixed with various concentrations of anti-PD-1 antibodies. Then normal human serum complements (A112, Quidel Corp., San Diego, CA, USA) diluted to certain concentrations were added. The cells were cultured at 37°C with 5% CO2 for 3 h. The cell lysis was assessed by the CellTiter-Glo cell viability kit. The target Ramos cells (American Type Culture Collection (ATCC), Manassas, VA, USA) were treated with rituximab (synthesized according to the C2B8 sequences described by the Genentech company in the patent US5736137) were used as the positive control. The percentage of target lysis was calculated as (1 – RFUsample/RFUtotal cells) * 100%.
Lou B., Wei H., Yang F., Wang S., Yang B., Zheng Y., Zhu J, & Yan S. (2021). Preclinical Characterization of GLS-010 (Zimberelimab), a Novel Fully Human Anti-PD-1 Therapeutic Monoclonal Antibody for Cancer. Frontiers in Oncology, 11, 736955.
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