Namalwa

Manufactured by American Type Culture Collection

Sourced in United States

The Namalwa is a cell line derived from a human Burkitt's lymphoma. It is a suspension-adapted cell line that is commonly used in research applications involving the study of viral infections, immunology, and cell biology.

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30 protocols using namalwa

Culturing Human Lymphoma Cell Lines

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The human Burkitt lymphoma cell lines Daudi (ATCC^® CCL-213™), Namalwa (ATCC^® CRL-1432™), Ramos (RA 1) (ATCC^® CRL-1596™), Raji (ATCC^® CCL-86™), the JeKo-1 (ATCC^® CRL-3006™) human mantle cell lymphoma cell line, and the RS4;11 (ATCC^® CRL-1432™) human acute lymphoblastic leukemia cell line were purchased from the ATCC (American Type Culture Collection, Manassas, VA, USA). All cells were maintained according to the ATCC instructions in an incubator at 37 °C with a humidified atmosphere of 5% CO₂ in air. Tissue culture flasks and dishes were purchased from Sarstedt, Inc. (Newton, NC, USA). Heat-inactivated fetal bovine serum (FBS) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Other cell culture media and reagents were purchased from Thermo Fisher Scientific (Waltham, MA, USA).

Chen L.M, & Chai K.X. (2023). Exosome-Mediated Activation of the Prostasin-Matriptase Serine Protease Cascade in B Lymphoma Cells. Cancers, 15(15), 3848.

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Comprehensive Cell Line Culturing Protocol

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All cell lines were cultured in a humidified incubator at 37°C with 5% CO₂. The following cell lines were obtained from ATCC: HEK 293 T, WI-38, WI-38 VA 13, Daudi, EB1, HS604T, HS616T, HuT 102, MC116, Namalwa, H1963, H196, H209, H526, H524, H82, H69, Raji, SW1271, and TO175T. DEL, L428, SU-DHL-10, WSU-DLCL2, Jurkat, DND41 and SR768 were purchased from DSMZ. A4/Fuk was obtained from JCRB Cell Line Bank. OCI-Ly3 was a kind gift from Dr. Mark Minden (University Health Network Toronto). The human lung fibroblast cell lines WI-38 and WI-38 VA13 were routinely cultured in EMEM supplemented with 10% Fetal Bovine Serum (FBS). SCLC cell lines H1963, H196, SW1271, H209, H526, H524, H82 and H69 were maintained in RPMI 1640 with 10% FBS. The lymphoma cell lines EB1, Daudi, Raji, A4/Fukuda, Jurkat, DND41, WSU-DLCL2, DEL, HUT102, Namalwa and L428 were cultured in RPMI 1640 with 10% FBS. MC116 and SU-DHL-10 cells were maintained in RPMI 1640 with 20% FBS. OCI-Ly3 cells were cultured in IMDM with 20% FBS. The SR786 cell line was cultured in RPMI 1640 with 15% FBS. HS604T cells were maintained in DMEM with 10%FBS and 2 mM Glutamine. HS616T and TO175T cell lines were cultured in DMEM with 10% FBS. HEK 293 T cells were maintained in DMEM with 10% FBS. All cell lines were maintained with a cocktail of penicillin and streptomycin (Gibco).

Zhou Z., Patel M., Ng N., Hsieh M.H., Orth A.P., Walker J.R., Batalov S., Harris J.L, & Liu J. (2014). Identification of synthetic lethality of PRKDC in MYC-dependent human cancers by pooled shRNA screening. BMC Cancer, 14, 944.

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Cell Line Characterization and Maintenance

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Ba/F3 (#HB-283), HeLa (#CCL-2), 293T (#CRL-1573), JeKo-1 (#CRL-3006), and Namalwa (#CRL-1432) were from ATCC, and Granta-519 (#ACC342) and Ri-1 (#ACC585) were from DSMZ. The 17p status of JeKo-1, Granta-519, Ri-1 and Namalwa have been confirmed by qPCR. All human lymphoma cell lines were cultured at 37 °C with 5% CO2 in RPMI-1640 medium supplemented with 10% v/v fetal bovine serum (FBS) and penicillin (100 units/ml)/streptomycin (0.1 mg/ml). Ba/F3 cells were maintained in RPMI-1640 medium supplemented with 10% v/v fetal bovine serum (FBS) and 2ng/ml IL3 (R&D Cat# 403-ML-050). All cell lines were routinely tested for mycoplasma by PCR. Experiments were performed within four weeks after fresh viable cells were thawed.

Chen M., Chen X., Li S., Pan X., Gong Y., Zheng J., Xu J., Zhao C., Zhang Q., Zhang S., Qi L., Wang Z., Shi K., Ding B.S., Xue Z., Chen L., Yang S., Wang Y., Niu T., Dai L., Lowe S.W., Chen C, & Liu Y. (2020). An epigenetic mechanism underlying chromosome 17p deletion-driven tumorigenesis. Cancer discovery, 11(1), 194-207.

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Diverse Cell Lines for Research

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The CA46, Raji, Namalwa, Ramos, Mino, Jeko-1, Maver-1, Z138, U266, NCI-H929, Jurkat, and HEK-293T cell lines were purchased (ATCC, Manassas, VA). The 207 and 697 cell lines were provided by Dr. P.D. Burrows (University of Alabama Birmingham). The HBL2, Toledo, BJAB, OCI-Ly19, SU-DHL-4, and Pfeiffer cell lines were provided by Dr. J. Tao (H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL). The U2932 cell line was provided by Dr. I.S. Lossos (Sylvester Comprehensive Cancer Center, Miami, FL) (37 (link)). Cells were cultured in Hyclone RPMI 1640 or DMEM media supplemented with 10% FBS (GE Healthcare Life Sciences, Pittsburgh, PA) and 1% penicillin-streptomycin and for NCI-H929 only 55 μM 2-mercapotoethanol.

Alexander L.E., Watters J., Reusch J.A., Maurin M., Nepon-Sixt B.S., Vrzalikova K., Alexandrow M.G., Murray P.G, & Wright K.L. (2017). Selective expression of the transcription elongation factor ELL3 in B cells prior to ELL2 drives proliferation and survival. Molecular immunology, 91, 8-16.

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Lymphoma and Leukemia Cell Lines Protocol

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Diffuse large B-cell lymphoma cell line SU-DHL-4, Burkitt lymphoma cell lines NAMALWA, RAMOS, RAJI and CA46, mantle cell lymphoma cell line JEKO-1, B lymphoblastic lymphoma cell line HMy2.CIR, B-ALL cell line NALM6, acute myeloid leukemia cell line MOLM13 were purchased from ATCC or DSMZ and preserved in our lab and all verified before being used in following experiments. All cell lines were cultured with RPMI 1640 medium containing 10% fetal bovine serum (FBS, Gibco, USA) in humidified 5% CO₂ at 37 °C. Primary human tumor cells were obtained by magnetic bead sorting from peripheral blood of 2 clinical patients who suffered relapse after CD19 CART treatment, 1 case of B-NHL and 1 case of B- ALL. We confirm that all methods were carried out in accordance with relevant guidelines and regulations or in accordance with the Declaration of Helsinki. We confirm that informed consent was obtained from all subjects and/or their legal guardian(s). And we confirm that all experimental protocols were approved by a named institutional and/or licensing committee.

Yang X., Yu Q., Xu H, & Zhou J. (2021). Upregulation of CD22 by Chidamide promotes CAR T cells functionality. Scientific Reports, 11, 20637.

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Targeting Epigenetic Regulators in Cancer

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Namalwa, Ramos, CA-46 and DAUDI cells were purchased from ATCC and maintained as instructed. Antibodies against BRD4 (#E2A7X), BRD2 (#5848), c-MYC (#D84C12), PARP (#46D11) were purchased from Cell Signaling Technology. Actin (#A5441) antibody was purchased from SigmaAldrich. BRD3 (#2088c3a) antibody was purchased from Santa Cruz Biotechnology. Secondary antibodies (#7074, #7076) were purchased from Cell Signaling Technology. MG132 (#M7449) was purchased from SigmaAldrich. Carfilzomib (#S2853) was purchased from Selleck. JQ1, OTX015, pomalidomide were synthesized according to methods published.

Lu J., Qian Y., Altieri M., Dong H., Wang J., Raina K., Hines J., Winkler J.D., Crew A.P., Coleman K, & Crews C.M. (2015). Hijacking the E3 Ubiquitin Ligase Cereblon to Efficiently Target BRD4. Chemistry & biology, 22(6), 755-763.

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Culturing Burkitt's Lymphoma Cell Lines

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Human burkitt's lymphoma cell lines Raji and NAMALWA were obtained from ATCC, USA, and maintained in RPMI 1640 medium supplemented with 10% or 20% FBS, 100 U/ml penicillin and 10 U/ml streptomycin at 37°C in a 5% CO₂, 95% humidified atmosphere. Luciferase‐expressing cell lines were generated through inducing lentiviral supernatant into the parental cell line according to reference.²²

Yang L., Li L., Meng M., Wang W., Yang S., Zhao Y., Wang R., Gao H., Tang W., Yang Y., Yang L., Liao L, & Hou Z. (2020). Evaluation of piggyBac‐mediated anti‐CD19 CAR‐T cells after ex vivo expansion with aAPCs or magnetic beads. Journal of Cellular and Molecular Medicine, 25(2), 686-700.

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Characterization of Burkitt Lymphoma Cell Lines

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Human Burkitt lymphoma cell lines Ramos, Daudi, NAMALWA, and Raji were purchased from ATCC. All cell lines were validated by the Characterized Cell Line Core Facility at MD Anderson using Short Tandem Repeat DNA profiling (STR fingerprinting). Ramos, Daudi, and Raji cell lines were maintained in RPMI 1640 with 10% FBS and 1% Penicillin-Streptomyocin. NAMALWA cells were maintained in RPMI 1640 with 7.5% FBS, 2 mM L-glutamine, 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, 10mM HEPES, and 1.0 mM sodium pyruvate. DMSO (Sigma) was used as a vehicle. Butyrolactone-3 (MB-3) inhibitor was purchased from Cayman Chemical and dissolved in DMSO.

Farria A.T., Mustachio L.M., Akdemir Z.H, & Dent S.Y. (2019). GCN5 HAT inhibition reduces human Burkitt lymphoma cell survival through reduction of MYC target gene expression and impeding BCR signaling pathways. Oncotarget, 10(56), 5847-5858.

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Genetically Modified Cell Lines for Research

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Nalm6 (ATCC CRL-3273), Namalwa (ATCC CRL-1432), Jurkat (ATCC TIB-152), HL-60 (ATCC CCL-240), HEK-293T (ATCC CRL-11268), and MiaPaCa2 (ATCC CRL-1420) cells were cultured as described elsewhere. Namalwa and Nalm6 were modified to express enhanced GFP (EGFP) and Nanoluciferase (NanoLuc) using the SELWP, as described previously.³³ (link) In addition, MiaPaCa2 cells were first transduced with SELWP and then with EhCD19-LVs to express EGFP-NanoLuc and human CD19.

Tristán-Manzano M., Maldonado-Pérez N., Justicia-Lirio P., Muñoz P., Cortijo-Gutiérrez M., Pavlovic K., Jiménez-Moreno R., Nogueras S., Carmona M.D., Sánchez-Hernández S., Aguilar-González A., Castella M., Juan M., Marañón C., Marchal J.A., Benabdellah K., Herrera C, & Martin F. (2022). Physiological lentiviral vectors for the generation of improved CAR-T cells. Molecular Therapy Oncolytics, 25, 335-349.

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CD70 Expression in Cell Lines

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Example 1

CD70 expression was measured in various cell lines. CD70 was found to be expressed, with a fragments/kilobase of exon/million reads mapped (FPKM) greater than 35, in 99% of the multiple myeloma tumor cell lines tested.

To further characterize the expression of CD70, CEM (ATCC), EoL-1 (Sigma), HL-60 (ATCC), KG-1a (ATCC), MV4; 11 (ATCC), NAMALWA (ATCC), Raji (ATCC), Toledo (ATCC), and U-937 (ATCC) cells were stained with an anti-CD70 antibody conjugated to PE (BD Pharmingen) in stain buffer (BD Pharmingen) for 30 minutes at 4° C. Cells were then washed and resuspended in stain buffer with propidium iodide (BD Pharmingen) prior to data acquisition. Samples were then acquired by flow cytometry and data analyzed (FIG. 2). CD70 expression was observed in the cell lines EoL-1, MV4; 11, NAMALWA, Raji, Toledo, and U-937 (FIG. 2), but not in the cell lines CEM, HL-60, and KG-1a (FIG. 2).

US11046775B2. CD70 binding molecules and methods of use thereof (2021-06-29). KITE PHARMA, INC. [US]. Inventors: Ruben Alvarez Rodriguez [US], Armen Mardiros [US].

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