XG human myeloma cell lines (HMCLs) were cultured in the presence of recombinant IL-6 as previously described [28 (link)]. XG7 Melphalan-resistant cell line was derived from the sensitive parental XG7 cells [29 (link)]. JJN3 was kindly provided by Dr Van Riet (Brussels, Belgium), JIM3 by Dr MacLennan (Birmingham, UK) and MM1S by Dr S. Rosen (Chicago, USA). AMO-1, LP1, L363, U266, OPM2, and SKMM2 were purchased from DSMZ (Braunsweig, Germany) and RPMI8226 from ATTC (Rockville, MD, USA). HMCLs were authenticated according to their short tandem repeat profiling and their gene expression profiling using Affymetrix U133 plus 2.0 microarrays deposited in the ArrayExpress public database under accession numbers E-TABM-937 and E-TABM-1088 [28 (link)].
Amo 1
Manufactured by Leibniz Institute DSMZ
Sourced in Germany, United States
The AMO-1 is a laboratory equipment designed for the analysis and manipulation of microorganisms. It provides a controlled environment for culturing and observing microbial samples. The core function of the AMO-1 is to facilitate the growth, study, and experimentation with various types of microorganisms.
Automatically generated - may contain errors
Lab products found in correlation
Skmm2, by Leibniz Institute DSMZ (9 mentions)
Nci h929, by Leibniz Institute DSMZ (7 mentions)
Rpmi8226, by American Type Culture Collection (6 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (6 mentions)
Opm 2, by Leibniz Institute DSMZ (5 mentions)
Molp 8, by Leibniz Institute DSMZ (4 mentions)
Rpmi 8226, by Leibniz Institute DSMZ (4 mentions)
Kms 12 bm, by Leibniz Institute DSMZ (4 mentions)
Hek293t, by American Type Culture Collection (3 mentions)
Mycoalert mycoplasma detection kit, by Lonza (3 mentions)
Lopra, by Leibniz Institute DSMZ (2 mentions)
Molp2, by Leibniz Institute DSMZ (2 mentions)
Hek293, by American Type Culture Collection (2 mentions)
Bt 474, by Leibniz Institute DSMZ (2 mentions)
Caki2, by Leibniz Institute DSMZ (2 mentions)
Mda mb 231, by Leibniz Institute DSMZ (2 mentions)
Nci h929, by American Type Culture Collection (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
U133 plus 2.0 microarrays, by Thermo Fisher Scientific (1 mentions)
23 protocols using amo 1
1
Culturing Human Myeloma Cell Lines
2
Characterization of Human Myeloma Cell Lines
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XGs human myeloma cell lines (HMCLs) were obtained as previously described [27 (link),28 (link)]. AMO-1, LP1, L363, OPM2, MOLP2, MOLP8, Lopra, and SKMM2 were purchased from DSMZ (Braunsweig, Germany), and RPMI8226 was purchased from ATCC (American Tissue Culture Collection, Rockville, MD, USA). JJN3 was kindly provided by Dr. Van Riet (Bruxelles, Belgium), and was provided MM1S by Dr. S. Rosen (Chicago, USA). HMCLs were authenticated according to their short tandem repeat profiling and their gene expression profiling using Affymetrix U133 plus 2.0 microarrays deposited in the ArrayExpress public database under accession numbers E-TABM-937 and E-TABM-1088.
3
Obtaining and Authenticating Human Myeloma Cell Lines
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XG HMCLs were obtained as previously described [18 (link)]. JJN3 was kindly provided by Dr. Van Riet (Brussels, Belgium), JIM3 by Dr. MacLennan (Birmingham, UK), and MM1S by Dr. S. Rosen (Chicago, USA). AMO-1, LP1, L363, U266, OPM2, and SKMM2 were purchased from DSMZ (Braunsweig, Germany) and RPMI8226 from ATTC (Rockville, MD, USA). All HMCLs derived in our laboratory were cultured in the presence of recombinant IL-6. HMCLs were authenticated according to their short tandem repeat profiling and their gene expression profiling using Affymetrix U133 plus 2.0 microarrays deposited in the ArrayExpress public database under accession numbers E-TABM-937 and E-TABM-1088 [18 (link)].
4
Cell Viability Assay for Drug Screening
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All cell lines were grown and treated in RPMI 1640 or Dulbecco’s modified Eagle medium (Sigma, St. Louis, MO) supplemented with 10% fetal bovine serum (Gibco) or 10% human serum (Sigma) and maintained in a humidified incubator at 5% CO2 and 37 °C. DLD-1, MV4-11, NCI-H929 (all ATCC, Manassas, VA, USA) and AMO-1 (DSMZ, Braunschweig, Germany) cells were plated 4000 cells per well in 384-well tissue culture plates and treated with compounds of interest for 24 h at 37 °C, 5% CO2 humidified incubator. Each concentration was tested in duplicate at least 3 separate times. Adherent cells were allowed to adhere overnight prior to treatment. Alternatively, AMO-1 or NCI-H929 cells were treated with ABBV-467 for the indicated contact times (1–24 h) at 37 °C in a 5% CO2 humidified incubator and washed 3 times with compound-free media prior to incubation for the remainder of a 24-h treatment period. Cell viability was subsequently determined using CellTiter-Glo (Promega, Madison, WI, USA) according to the manufacturer’s instructions. Half maximal effective concentration (EC50) and 90% of the maximal effective concentration (EC90) values were calculated from the resulting concentration-response curves using non-linear regression analysis.
5
Myeloma Cell Lines and Controls
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XGs human myeloma cell lines (HMCLs) were obtained as previously described 8 (link). AMO-1, LP1, L363, OPM2, MOLP2, MOLP8, Lopra and SKMM2 were purchased from DSMZ (Braunsweig, Germany) and RPMI8226 from ATCC (American Tissue Culture Collection, Rockville, MD, USA). JJN3 was kindly provided by Dr. Van Riet (Bruxelles, Belgium) and MM1S by Dr. S. Rosen (Chicago, USA). HMCLs were authenticated according to their short tandem repeat profiling and their gene expression profiling using Affymetrix U133 plus 2.0 microarrays deposited in the ArrayExpress public database under accession numbers E-TABM-937 and E-TABM-1088. HMCLs characteristics, obtained from previously published analysis results 8 (link), are available in Table S1 . EBV-immortalized B-cells from 8 different patients have been used as control cells. The patients are those from whom the XG1, XG3, XG5, XG10, XG13, XG14, XG16 and XG19 cell lines were generated.
6
Human Myeloma Cell Lines and Primary Samples
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XG human myeloma cell lines were obtained as previously described 16 (link). AMO-1, OPM2 and SKMM2 were purchased from DSMZ (Braunschweig, Germany) and RPMI8226 from ATCC (Manassas, USA). JJN3 was kindly provided by Dr.Van Riet (Brussels, Belgium), and KMS-12-BM by Dr Otsuki (Okayama, Japan). HMCLs characteristics are available in Table 1 .
Bone marrow samples were collected after patient's written informed consent in accordance with the Declaration of Helsinki and institutional research board approval from Montpellier University Hospital. Bone marrow was collected from 69 patients at diagnosis and 28 patients at relapse, this cohort was called Montpellier cohort. MM cells of patients were purified using anti-CD138 MACS microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany). We also used RNA sequencing data of 674 newly diagnosed MM patients with longitudinal follow-up from the Multiple Myeloma Research Foundation CoMMpass trial (NCT01454297; version IA11a), termed in the following CoMMpass cohort.
Normal plasma cells were generated using a 3-step in vitro model starting from purified memory B cells from 3 different healthy donors as reported 17 (link),18 (link).
Bone marrow samples were collected after patient's written informed consent in accordance with the Declaration of Helsinki and institutional research board approval from Montpellier University Hospital. Bone marrow was collected from 69 patients at diagnosis and 28 patients at relapse, this cohort was called Montpellier cohort. MM cells of patients were purified using anti-CD138 MACS microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany). We also used RNA sequencing data of 674 newly diagnosed MM patients with longitudinal follow-up from the Multiple Myeloma Research Foundation CoMMpass trial (NCT01454297; version IA11a), termed in the following CoMMpass cohort.
Normal plasma cells were generated using a 3-step in vitro model starting from purified memory B cells from 3 different healthy donors as reported 17 (link),18 (link).
7
Profiling Multiple Myeloma Drug Resistance
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Across the study, following cell lines were used: Multiple myeloma cell lines AMO-1 (DSMZ, German Collection of Microorganisms and Cell Cultures GmbH) and its derivatives resistant to proteasome inibitors bortezomib: AMO-BTZ and carfilzomib: AMO-CFZ. Further following cell lines were used: MDA-MB-231 (DSMZ), BT-474 (DSMZ), U-2 OS (ATCC) K562 (ATCC), HeLa (ATCC), HEK293 (ATCC), HEK293T (ATCC), and Caki2 (DSMZ). The cells were authenticated by STR-typing and routinely tested for Mycoplasma contamination (MycoAlert Mycoplasma Detection Kit). For detailed information about cell lines maintenance, see Supplementary Methods. For the complete list of chemicals used across the study, see Supplementary Table S1 .
8
Cultivation of Multiple Myeloma Cell Lines
9
Culturing Multiple Myeloma Cell Lines
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NCI-H929, RPMI-8226, AMO-1, and AMO-BZB MM cell lines were purchased from DSMZ, certified authentication performed by short tandem repeat DNA typing, and were cultured in RPMI-1640 media containing 10% fetal bovine serum (FBS) (Corning, Tewksbury MA, USA), 2 μmol/L glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin (Sigma Aldrich, St Louis, MO, USA) at 37 °C in a 5% CO2 atmosphere. Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll-hypaque (Lonza Group, Basel, Switzerland) following the manufacturer’s protocol.
10
Characterization of Human Myeloma Cell Lines
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All (HMCLs) were previously characterized.12,22 (link) The HMCLs BCN; NAN1, −3, −7, −8 and −9; SBN; and XG1, −2, −5, −6, −7 and −11 were derived in Nantes or Montpellier laboratories in the presence of interleukin-6 (IL-6). KMS11, KMS12-PE and KMM1 HMCLs were kindly provided by Dr. Otsuki (Kurashiki, Japan), JJN3 was kindly provided by Dr. Van Riet (Brussels, Belgium), JIM3 was kindly provided by Dr. MacLennan (Birmingham, UK), Karpas 620 was kindly provided by Dr. Karpas (Cambridge, UK) and MM1S was kindly provided by Dr. Rosen (Chicago, IL). AMO1, LP1, L363, NCI-H929, SKMM2, U266 and OPM2 were purchased from DSMZ (Braunsweig, Germany). All HMCLs were cultured in RPMI1640 supplemented with 5% fetal calf serum (FCS). Cell cultures from BCN, NAN, SBN and XG cells were additionally supplemented with 3 ng/mL IL-6. Blood or bone marrow samples from patients were collected after informed consent at the Department of Hematology, University Hospital of Nantes. Experiments with primary myeloma cells were carried out using unpurified or purified (CD138+) myeloma cells, depending on the myeloma infiltration and the total number of cells. Chromosome abnormalities were assessed by fluorescence in situ hybridization (FISH).
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