The following cell lines (Kasumi-1, NB4, NALM-6, Jurkat, K562, Daudi, IM-9, RPMI8226, NCI-H929, HeLa, and RK13) were obtained from ATCC between 2004 and 2013. FL-218 was obtained from the Japanese Collection of Research Bioresources Cell Bank in 2011. Normal human skin fibroblasts (NB1RGB) were obtained from the RIKEN BioResource Center in 2014. All cell lines were passaged for less than 6 months. No further authentication was done for these cell lines in the past 6 months. Bone marrow samples were collected from patients admitted to the Japanese Red Cross Medical Center between February 2014 and December 2014; written informed consent was obtained from all patients prior to collection. All relevant study-related protocols were approved by the Institutional Review Board of the Japanese Red Cross Medical Center and the Institute of Medical Science, The University of Tokyo. Cells were grown in appropriate culture medium supplemented with 10% fetal bovine serum (JRH Biosciences), 100 U/mL penicillin (Wako), and 100 μg/mL streptomycin (Wako). Cell lines derived from hematological malignancies and patient bone marrow mononuclear cells were grown in RPMI 1640 medium (Wako). HeLa cells and NB1RGB cells were grown in minimum essential medium (MEM), alpha modification (Wako). RK13 cells were grown in standard MEM (Wako).
Nci h929
Manufactured by American Type Culture Collection
Sourced in United States
NCI-H929 is a cell line derived from a human multiple myeloma. It is maintained in culture and available for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi8226, by American Type Culture Collection (55 mentions)
Mm 1s, by American Type Culture Collection (42 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (33 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (26 mentions)
Penicillin, by Thermo Fisher Scientific (20 mentions)
Streptomycin, by Thermo Fisher Scientific (20 mentions)
Rpmi 1640, by Thermo Fisher Scientific (10 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (9 mentions)
Fetal bovine serum (fbs), by Merck Group (8 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (7 mentions)
L glutamine, by Thermo Fisher Scientific (6 mentions)
Mm 1r, by American Type Culture Collection (6 mentions)
Arh 77, by American Type Culture Collection (5 mentions)
Hek293t, by American Type Culture Collection (5 mentions)
Molp 8, by Leibniz Institute DSMZ (5 mentions)
Kms 12 pe, by Leibniz Institute DSMZ (5 mentions)
Nci h929, by Thermo Fisher Scientific (5 mentions)
Mycoalert mycoplasma detection kit, by Lonza (4 mentions)
Jurkat, by American Type Culture Collection (4 mentions)
Kms 11, by Japanese Collection of Research Bioresources Cell Bank (4 mentions)
100 protocols using nci h929
1
Cell Line Origin, Culture, and Consent
2
Characterization of Multiple Myeloma Cell Lines
3
Cell Culture Conditions for Myeloma and CLL
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Cells were cultured in incubators containing
5% CO2 and maintained at 37 °C. RPMI-8226 and NCI-H929
human MM cell lines (purchased from ATCC), 5TGM1 mouse MM cell line
(a gift from Dr. Lori A. Hazlehurst at the West Virginia University,
Morgantown, WV), MEC2 and WaC3 human CLL cell lines (gifts from Dr.
Javier A. Pinilla-Ibarz at the Moffitt Cancer Center, Tampa, FL),
and primary B cells purified from spleens of mice were grown in RPMI
1640 media (Gibco) supplemented with heat-inactivated fetal bovine
serum (FBS, 10%),l -glutamine (2 mM), sodium pyruvate (1
mM), nonessential amino acids (0.1 mM), β-mercaptoethanol (β-ME;
0.1 mM), penicillin G sodium (100 U/mL), and streptomycin sulfate
(100 μg/mL). The J558 mouse myeloma cell line (purchased from
ATCC) was cultured in DMEM media (Gibco) and 10% heat-inactivated
horse serum together with the abovementioned supplemental nutrients.
Human embryonic kidney 293 T cells (purchased from ATCC) and mouse
hepatoma HEPA 1–6 cell line (purchased from ATCC) were cultured
in DMEM media with 10% heat-inactivated FBS and the same supplemental
nutrients.
5% CO2 and maintained at 37 °C. RPMI-8226 and NCI-H929
human MM cell lines (purchased from ATCC), 5TGM1 mouse MM cell line
(a gift from Dr. Lori A. Hazlehurst at the West Virginia University,
Morgantown, WV), MEC2 and WaC3 human CLL cell lines (gifts from Dr.
Javier A. Pinilla-Ibarz at the Moffitt Cancer Center, Tampa, FL),
and primary B cells purified from spleens of mice were grown in RPMI
1640 media (Gibco) supplemented with heat-inactivated fetal bovine
serum (FBS, 10%),
mM), nonessential amino acids (0.1 mM), β-mercaptoethanol (β-ME;
0.1 mM), penicillin G sodium (100 U/mL), and streptomycin sulfate
(100 μg/mL). The J558 mouse myeloma cell line (purchased from
ATCC) was cultured in DMEM media (Gibco) and 10% heat-inactivated
horse serum together with the abovementioned supplemental nutrients.
Human embryonic kidney 293 T cells (purchased from ATCC) and mouse
hepatoma HEPA 1–6 cell line (purchased from ATCC) were cultured
in DMEM media with 10% heat-inactivated FBS and the same supplemental
nutrients.
4
Generating Bortezomib-Resistant Myeloma Cell Lines
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The human multiple myeloma cell lines MM.1S, MM.1R, OPM1, RPMI8226/Dox, and NCIH929 were purchased from ATCC Company. Cells were grown in suspension in RMPI1640 medium supplemented with 10% fetal bovine serum, 1% (v/v) penicillin, and 100 μg/mL streptomycin. Cells were maintained at 37 °C in a 5% CO2 atmosphere with a proper humidity.
To generate bortezomib-resistant myeloma cell lines, bortezomib (BTZ) was added to the multiple myeloma cell culture medium starting at 0.03 nM. The culture medium was replaced with bortezomib containing medium twice weekly and the myeloma cells were cultured for 2–4 weeks until the cells survived and became resistant to that concentration of bortezomib. The bortezomib concentration was then increased by doubling the previous concentration until BTZ reached at a final concentration of up to 8.4 nM by the end of the second year. Myeloma cell lines were maintained on BTZ containing medium until 1 week before experiments.
To generate bortezomib-resistant myeloma cell lines, bortezomib (BTZ) was added to the multiple myeloma cell culture medium starting at 0.03 nM. The culture medium was replaced with bortezomib containing medium twice weekly and the myeloma cells were cultured for 2–4 weeks until the cells survived and became resistant to that concentration of bortezomib. The bortezomib concentration was then increased by doubling the previous concentration until BTZ reached at a final concentration of up to 8.4 nM by the end of the second year. Myeloma cell lines were maintained on BTZ containing medium until 1 week before experiments.
5
Establishing Cell Culture Conditions
6
Establishment and Characterization of Multiple Myeloma Cell Lines
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Human MM cell lines MM.1S, RPMI-8226, OPM-2, U266 and NCI-H929 were respectively purchased from ATCC and DSMZ, and they were cultured with RPMI-1640 medium (Gibco) supplemented with 10% fetal bovine serum (FBS, BI) at 37℃ in 5% CO2. All cell lines have been tested negative for contamination with mycoplasma. MM.1S-mCherry.ffLuc, RPMI-8226-mCherry.ffLuc, BCMA-GFP-Hela and CD47-GFP-Hela cells were prepared through lentivirus infection. pcDNA3.1 vector (cat# V79020, Invitrogen) was used for eukaryotic expression of the recombinant proteins, pMECS vector was used to construct phage library, LentiGuide-Puro (cat# 52963) and pSLCAR-CD19-BBz (cat# 135992) vectors were purchased from Addgene and used to prepare lentivirus.
7
Culturing Multiple Myeloma Cell Lines
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Multiple myeloma cell lines MM.1S, OPM2, NCI-H929, L363, LP1, RPMI-8226, AMO-1, INA-6, JJN3, and KMS12BM were obtained from ATCC (Manassas, Virginia, USA) and the German Collection of Microorganisms and Cell Cultures GmbH (DSMZ, Braunschweig, Germany), and maintained in RPMI-1640 medium (Merck KGaA, Darmstadt, Germany) containing 10% fetal bovine serum (FBS) and supplemented with 1% penicillin/streptomycin and 1% l -glutamine. INA-6 cells were supplemented with IL-6. Cells were maintained at 37 °C with 5% CO2 in humidified atmosphere.
8
Culturing Human Multiple Myeloma Cell Lines
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The L363, MM.1S, NCI-H929, RPMI8226, and U266 human MM cell lines, as well as the HEK293T human embryonic kidney cell line, were purchased from ATCC. MOLP-8 cells were purchased from DSMZ. The KMM-1, KMS-11, KMS-12 PE, KMS-21 BM, KMS-26, KMS-27, and KMS-34 cell lines were obtained from JCRB (National Institute of Health Sciences). OPM2 cells were provided by Dr. Naoki Hosen (Osaka University). The MM.1R cell line was obtained from Dr. Steven Rosen. All MM cell lines were cultured in 5% CO2 at 37°C in RPMI1640 medium (Thermo Fisher Scientific) containing 10% FBS (Sigma-Aldrich), 2 mmol/L L-glutamine, 100 U/mL penicillin, and 100 mg/mL streptomycin (Invitrogen). HEK293T cells were cultured in DMEM (Thermo Fisher Scientific) containing 10% FBS, 100 U/mL penicillin, and 100 mg/mL streptomycin. Cell lines were used within 3 months after thawing. Mycoplasma contamination was checked for using the MycoAlert Mycoplasma Detection Kit (Lonza).
9
Evaluating IFNα2b Effects on MM Cell Viability
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Viability of MM cell lines cultured with and without 10,000 IU/ml IFNα2b (US Biological, Salem, OR) was determined using the CellTiter-Glo® luminescent assay (Promega, Madison, WI). MM cell lines included ARH-77 (ATCC® CCL-155™), RPMI8226 (ATCC® CRL-1621™), NCI-H929 (ATCC® CRL-9068™), U266 (ATCC® TIB-196™) and ARP-1 (Myeloma Institute, University of Arkansas, Little Rock, AK). IFNα activity on CD38-negative reporter cells (iLite™ cell line) was determined using the iLite™ Cell Assay following a modified manufacturer’s protocol (PBL Assay Science, Piscataway, NJ).
10
Cell Line Collection and Preparation
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