Nci h929

Manufactured by Thermo Fisher Scientific

Sourced in United States

The NCI-H929 is a human multiple myeloma cell line. It is a commonly used in vitro model for research on multiple myeloma.

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9 protocols using nci h929

Circ-SMARCA5 Expression in Multiple Myeloma

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In order to further explore the role of Circ-SMARCA5 in MM etiology, cells experiments were subsequently performed. Wild type MM cell lines (NCI-H929, RPMI8226, U226, OPM2 and JJN3) were purchased from Chinese Academy of Sciences (Beijing, China) or kindly given by Shanghai Jiaotong University (Shanghai, China), and normal plasma cells were isolated from bone marrow of healthy donors using CD138-coated magnetic beads (Miltenyi Biotec, Germany) as normal control. NCI-H929, RPMI8226, U226 and OPM2 cells were cultured in 90% RPMI-1640 medium (Gibco, USA) (containing 1.5 g/L NaHCO₃, 2.5 g/L glucose and 0.11 g/L sodium pyruvate) supplemented with 10% fetal bovine serum (FBS) (Gibco, USA); JJN3 cells were cultured in 40% DMEM medium (Gibco, USA) and 40% IMEM medium (Gibco, USA) supplemented with 20% FBS (Gibco, USA). All cells were incubated at 37 °C under 95% air and 5% CO₂ condition.
Measurement of Circ-SMARCA5 in MM cell lines and normal plasma cells.
The qPCR was performed to detect the expression of Circ-SMARCA5 in wild-type MM cell lines: NCI-H929, RPMI8226, U226, OPM2 and JJN3, and normal plasma cells.

Liu H., Wu Y., Wang S., Jiang J., Zhang C., Jiang Y., Wang X., Hong L, & Huang H. (2019). Circ-SMARCA5 suppresses progression of multiple myeloma by targeting miR-767-5p. BMC Cancer, 19, 937.

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SIRT2 Expression in Multiple Myeloma Cell Lines

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Human MM cell lines, KMS-28BM, U266, RPMI-8226 and NCI-H929, were purchased from the American Type Culture Collection. All cells were cultured in RPMI-1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% (KMS-28BM, RPMI-8226 and NCI-H929) or 15% fetal bovine serum (U266) (Gibco; Thermo Fisher Scientific, Inc.) and 1% penicillin and streptomycin at 37°C in a humidified incubator containing 5% CO₂. The plasma cells were isolated from bone marrow mononuclear cells, which were derived from the bone marrow samples of patients with MM and healthy bone marrow donors. Briefly, after collection, the bone marrow samples were processed with gradient density centrifugation (865 × g at 37°C for 20 min) for separating the bone marrow mononuclear cells; subsequently, the separated bone marrow mononuclear cells were purified using CD138-coated magnetic beads (Miltenyi Biotec GmbH) to obtain plasma cells (22 (link),23 (link)). The plasma cells were then stored in liquid nitrogen for further analysis. After incubation at 37°C for 24 h, SIRT2 expression in MM cell lines and normal plasma cells (from healthy bone marrow donors that were used as the control group) was determined by RT-qPCR and western blot analysis.

Ding T, & Hao J. (2021). Sirtuin 2 knockdown inhibits cell proliferation and RAS/ERK signaling, and promotes cell apoptosis and cell cycle arrest in multiple myeloma. Molecular Medicine Reports, 24(5), 760.

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Culturing Multiple Myeloma Cell Lines

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The human MM cell lines U266, MM.1S, RPMI8226, and NCI-H929 were purchased from the American Type Culture Collection (Manassas, VA, USA). The KMS-11, KMS-12-BM, KMS-12-PE, KMS-28-BM, and KMS-28-PE cell lines were purchased from the Japanese Collection of Research Bioresources Cell Bank (JCRB, Osaka, Japan). All MM cell lines except NCI-H929 were cultured in RPMI1640 (GIBCO, Thermo Fisher, Grand Island, NY, USA) containing 10% fetal bovine serum (GIBCO, Thermo Fisher) and 1% penicillin/streptomycin (P/S) (GIBCO, Thermo Fisher).

Lee K.J., Lee K.H., Yoon K.A., Sohn J.Y., Lee E., Lee H., Eom H.S, & Kong S.Y. (2017). Chromothripsis in Treatment Resistance in Multiple Myeloma. Genomics & Informatics, 15(3), 87-97.

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Culturing Human Multiple Myeloma Cell Lines

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RPMI-8226, NCI-H929, U266 and ARH-77 human MM cell lines were purchased from American Type Culture Collection (Manassas, VA, USA). The RPMI-8226, NCI-H929, U266 and ARH-77 cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) and 100 U/ml penicillin/streptomycin (Beyotime Institute of Biotechnology, Haimen, China). The cells were incubated in a humidified incubator with 5% CO₂ at 37°C.

Mei H., Xiang Y., Mei H., Fang B., Wang Q., Cao D., Hu Y, & Guo T. (2018). Pterostilbene inhibits nutrient metabolism and induces apoptosis through AMPK activation in multiple myeloma cells. International Journal of Molecular Medicine, 42(5), 2676-2688.

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Culturing Human Multiple Myeloma Cell Lines

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Human MM cell lines (including NCI-H929, U266, LP-1 and RPMI-8226) were all purchased from Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbH (DSMZ, Germany). The 293T cells were purchased from American Type Culture Collection (ATCC, USA). The NCI-H929 cells, U266 cells and RPMI-8226 cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 Medium (Gibco, USA) containing 10% fetal bovine serum (FBS) (Gibco, USA). The LP-1 cells were cultured in Iscove’s Modified Dulbecco’s Medium (IMDM) containing 10% FBS (Gibco, USA), and 293T cells were cultured in Dulbecco’s Modified Eagle’s Medium (Gibco, USA) containing 10% FBS (Gibco, USA). All cells were maintained in a humidity atmosphere of 5% CO₂ at 37°C. Human normal bone marrow mononuclear cells (BMMCs) were purified with CD138-coated magnetic beads (Miltenyi Biotec, Germany) from bone marrow samples donated by health donors. This study was approved by Shanghai Jing’an District Zhabei Central Hospital and conducted by following the guideline of Ethical Guidelines for Human Genome/Gene Research issued by the Chinese Government. Written informed consents were provided by all participants before enrollment.

Zhou F., Wang D., Zhou N., Chen H., Shi H., Peng R., Wei W, & Wu L. (2021). Circular RNA Protein Tyrosine Kinase 2 Promotes Cell Proliferation, Migration and Suppresses Apoptosis via Activating MicroRNA-638 Mediated MEK/ERK, WNT/β-Catenin Signaling Pathways in Multiple Myeloma. Frontiers in Oncology, 11, 648189.

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Cell Culture Maintenance of Bone Marrow and Myeloma Cells

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Bone marrow stromal cell line HS-5 and the human MM cell lines RPMI 8226 and NCI-H929 were purchased from Cell Library, China Academy of Science. The cell lines NCI-H929 and RPMI 8226 were grown in RPMI 1640 (Gibco BRL, Grand Island, NY, USA) and the HS-5 was cultured in Dulbecco's Modified Eagle's Medium (DMEM; Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (Gibco BRL). The cells were cultured in a cell incubator containing 5% CO 2 under the saturated humidity at 37°C.

Shen Y., Liu H., Gu S., Wei Z, & Liu H. (2017). The role of Capon in multiple myeloma. Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine, 39(7).

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Establishing Myeloma Cell Lines and Mouse Models

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Human myeloma cell lines (NCI-H929, U266, MM1.S and RPMI-8226), human embryonic kidney epithelial cell (HEK-293T) and human cervical cancer cell HeLa were acquired from the American Type Culture Collection (ATCC; USA). The human myeloma cell line KMS11 and the ARD cell line was kindly donated from Prof. Yuhuan Zheng (West China hospital, Sichuan University). The ADR, KMS11, NCI-H929, U266, MM1.S and RPMI-8226 cells were cultured in RPMI-1640 medium (Thermo Fisher Scientific). HEK-293T cells were maintained in DMEM (Gibco; USA). Cell line culture media was supplemented with 10% fetal bovine serum (Gibco; USA), penicillin and streptomycin (1.0 mmol/L, HyClone; USA). Cell lines were grown at 37 °C in a humid incubator with 5% CO₂.
Female NSG and B-NDG mice were obtained from Nanjing University’s Model Animal Resource Information Platform and maintained in a pathogen-free setting at the State Key Laboratory of Biotherapy, Sichuan University. All the experimental procedures were performed in compliance with the Guide for the Care and Use of Laboratory Animals of Sichuan University and approved by the Animal Care and Use Committee of Sichuan University (reference numbers: 2018–061).

Yu Z., Li H., Lu Q., Zhang Z., Tong A, & Niu T. (2024). Fc receptor-like 5 (FCRL5)-directed CAR-T cells exhibit antitumor activity against multiple myeloma. Signal Transduction and Targeted Therapy, 9, 16.

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Culturing Multiple Myeloma Cell Lines and Normal Plasma Cells

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Three MM cell lines (NCI-H929, MM.1S, and U266) were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). Normal plasma cells (nPCs) were sorted from bone marrow of healthy volunteers by the FACS Calibur flow cytometer cytometry (BD Biosciences, San Jose, CA, USA) via the following four surface markers: CD45-PB, CD56-PE, CD38-FITC, and CD19-PC7. Immunophenotype of nPCs was CD45+/dimCD38+CD19+CD56−. NCI-H929 cells were grown in the RPMI-1640 medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 2-mercaptoethanol (0.05 mM; Sigma-Aldrich, St Louis, MO, USA) and 10% fetal bovine serum (FBS; Thermo Fisher Scientific). MM.1S, U266 and nPCs cells were cultured in the RPMI-1640 medium (Thermo Fisher Scientific) containing 10% FBS. All the cells were maintained in a humidified air of 5% carbon dioxide (CO₂) at 37°C.

Zhang Y., Zhao D., Li S., Xiao M., Zhou H., Yang S., Hao Y, & Dong S. (2020). Long non-coding RNA TUG1 knockdown hinders the tumorigenesis of multiple myeloma by regulating the microRNA-34a-5p/NOTCH1 signaling pathway. Open Life Sciences, 15(1), 284-295.

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Culturing and Sourcing Multiple Myeloma Cell Lines

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Cell lines L363, OPM-2, MM1.S, RPMI-8226, and SU-DHL-2 were cultured in RPMI 1640 GlutaMAX HEPES culture medium (Life Technologies) supplemented with 10% fetal bovine serum (FBS, Biowest) and 100 µg/ml penicillin–streptomycin (Gibco/Life Technologies). UM9 was cultured with 20% FBS, and NCI-H929 with 20% FBS, 1 mM sodium pyruvate (Thermo Fisher), and 50 µM β-mercaptoethanol (Life Technologies). OCI-Ly1, OCI-Ly3, OCI-Ly7, and OCI-Ly10 were cultured in Iscove’s modified Dulbecco’s medium (Life Technologies) supplemented with 20% FBS and 100 µg/ml penicillin–streptomycin. Cell lines L363, RPMI-8226, OPM-2, OCI-Ly3, OCI-Ly7, OCI-Ly1 and NCI-H929 were purchased from DSMZ, MM1.S, OCI-Ly10, and SU-DHL-2 were purchased from ATCC. Cell line UM9 was derived from an MM patient at our UMCU hospital). All cell lines were tested negative for mycoplasma before and during the experiments. Primary MM cell samples were derived from patients diagnosed at the Academic Medical Center, Amsterdam, The Netherlands. This study was conducted and approved by the AMC Medical Committee on Human Experimentation. Informed consent was obtained in accordance with the Declaration of Helsinki.

Slomp A., Moesbergen L.M., Eldering E., Kersten M.J., Minnema M.C, & Peperzak V. (2021). Phosphatase PP2A enhances MCL-1 protein half-life in multiple myeloma cells. Cell Death & Disease, 12(3), 229.

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