Mm 1s

Manufactured by American Type Culture Collection

Sourced in United States, Germany

The MM.1S is a cell culture incubator designed for maintaining optimal conditions for cell growth and proliferation. It provides a controlled environment with precise temperature, humidity, and gas composition regulation. The MM.1S is a core laboratory equipment piece suitable for a variety of cell culture applications.

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Lab products found in correlation

231 protocols using mm 1s

Transduction and Isolation of Human Myeloma, Mesenchymal, and Endothelial Cells

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Human myeloma cell lines L363 and MM1S were purchased from the American Type Culture Collection and were retro-virally transduced (L363-GFP and MM1S-mCherry) as described previously.25 (link),26 (link) Human MSCs were obtained from the acetabular BM of patients undergoing hip replacement surgery as described previously.23 (link) Primary EPCs were obtained from umbilical cord blood collected from full-term pregnancies as described previously.27 (link) Human BM was obtained from the spina iliaca posterior superior of myeloma patients. The CD138⁺ cell population was isolated from the mononuclear cells of the myeloma BM by microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany) according to manufacturer’s protocol, and used immediately in the coculture system. Used protocols were approved by the local ethics committee of the University Medical Center Utrecht in accordance with the Declaration of Helsinki; all samples were obtained after written informed consent. Culture conditions can be found in the Materials and methods in the Supplementary material.

Braham M.V., Deshantri A.K., Minnema M.C., Öner F.C., Schiffelers R.M., Fens M.H, & Alblas J. (2018). Liposomal drug delivery in an in vitro 3D bone marrow model for multiple myeloma. International Journal of Nanomedicine, 13, 8105-8118.

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Culturing Multiple Myeloma and Endothelial Cells

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The MM cell lines, MM.1S and H929, were purchased from the American Type Culture Collection (ATCC, Rockville, MD), OPM-2 and green fluorescent protein-labeled and luciferase-transfected MM.1S (MM.1S-GFP-Luc) were a kind gift from Dr. Irene Ghobrial (Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA). Human umbilical vein endothelial cells (HUVECs) were purchased from Angio-Proteomie (Boston, MA). Human samples for this study were collected under informed consent, in concordance with Washington University Institutional Review Board (IRB) approval (IRB protocol number 201102270). MM cell lines were cultured in RPMI-1640 media (Corning, Tewksbury, MA) supplemented with 10% fetal bovine serum (FBS; Gibco, Life Technologies, Grand Island, NY), 2 mmol/L L-glutamine, 100 μg/mL penicillin, and 100 μg/mL streptomycin (Corning CellGro). HUVECs were cultured in Endothelial Growth Medium (EGM, Angio-Proteomie, Boston, MA) supplemented with endothelial growth supplements (including 10% FBS, recombinant growth factors, and 1% penicillin and streptomycin). All cells were cultured at 37 °C and in 5% CO₂ in a NuAire water jacket incubator (Plymouth, MN).

Federico C., Alhallak K., Sun J., Duncan K., Azab F., Sudlow G.P., de la Puente P., Muz B., Kapoor V., Zhang L., Yuan F., Markovic M., Kotsybar J., Wasden K., Guenthner N., Gurley S., King J., Kohnen D., Salama N.N., Thotala D., Hallahan D.E., Vij R., DiPersio J.F., Achilefu S, & Azab A.K. (2020). Tumor microenvironment-targeted nanoparticles loaded with bortezomib and ROCK inhibitor improve efficacy in multiple myeloma. Nature Communications, 11, 6037.

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Cell Line Authentication and Characterization

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MC/CAR cells were purchased in 2013 from ATCC (Manassas, VA), MM.1S and U266B cells were purchased from ATCC in 2016. OPM2 cells were purchased from The Leibniz Institute DSMZ in 2013 (Braunschweig, Germany). ARP-1 and ARD cells were obtained from Dr. Jonathan J. Keats in 2016 (University of Arkansas for Medical Sciences/Translational Genomics Research Institute, Phoenix, AZ). All cell lines were maintained in RPMI, high glucose medium (Corning, Corning, NY) supplemented with 20% heat-inactivated fetal bovine serum (Thermo Scientific, Grand Island, NY), 2mM GlutaMAX (Thermo Scientific), and 1× penicillin/streptomycin (Corning).
Cells used in the experiments described below were passaged fewer than 5 times since receipt from the vendors. MM.1S and U266B1 cells tested negative for Mycoplasma pulmonis on June 13, 2016; ARP-1, ARD and MC/CAR cells tested negative for Mycoplasma pulmonis on November 30, 2015. All cell lines were authenticated by short tandem repeat (STR) profiling at ATCC in November 2017. Two cell lines (ARD and ARP-1) were not part of the ATCC or DSMZ STR database. STR analysis of late passage of ARD and ARP-1 cells matched early passage cells obtained from Dr. Jonathan J. Keats.

Abrahams C.L., Li X., Embry M., Yu A., Krimm S., Krueger S., Greenland N.Y., Wen K.W., Jones C., DeAlmeida V., Solis W.A., Matheny S., Kline T., Yam A.Y., Stafford R., Wiita A.P., Hallam T., Lupher M, & Molina A. (2018). Targeting CD74 in multiple myeloma with the novel, site-specific antibody-drug conjugate STRO-001. Oncotarget, 9(102), 37700-37714.

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Cell Culture Conditions for HEK293T, HeLa, and Multiple Myeloma Cell Lines

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HEK293T (ATCC: CRL-3216), CRBN^−/− HEK293FT (kindly provided by WG Kaelin) and HeLa (ATCC: CCL-2) cells were grown in Dulbecco’s modified Eagle’s medium (DMEM; GIBCO) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin. The human multiple myeloma cell lines MM1.S (ATCC: CRL-2974), MM1.S CRBN^−/− (independent clones T11 and T21; both kind gifts of WG Kaelin), U266 (DSMZ: ACC-9), KMS 12BM (DSMZ: ACC-551), RPMI 8226 (DSMZ: ACC-402), JJN3 (DSMZ: ACC-541) were cultured in RPMI-1640 (GIBCO) with 10% heat-inactivated FBS and 1% penicillin-streptomycin. All cells tested mycoplasma negative by a PCR detection method.

Heider M., Eichner R., Stroh J., Morath V., Kuisl A., Zecha J., Lawatscheck J., Baek K., Garz A.K., Rudelius M., Deuschle F.C., Keller U., Lemeer S., Verbeek M., Götze K.S., Skerra A., Weber W.A., Buchner J., Schulman B.A., Kuster B., Fernández-Sáiz V, & Bassermann F. (2021). The IMiD target CRBN determines HSP90 activity toward transmembrane proteins essential in multiple myeloma. Molecular Cell, 81(6), 1170-1186.e10.

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Cell Culture Techniques for Various Cell Lines

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HeLa and HEK293T cells (ATCC) were grown in Dulbecco’s Modified Eagle’s Medium (DMEM, + 4.5 g/L D-glucose, + L-Glutamine, Sigma) supplemented with 10% FBS. HLF cells (Broad Institute CCLE) were grown in DMEM with 5% FBS. CUTLL1 (generously provided by J. Aster, Brigham and Women’s Hospital) and MM1.S (ATCC) cells were grown in RPMI 1640 (+ L-Glutamine, Sigma) supplemented with 10% FBS. Cells were maintained at 37°C and 5% CO₂. HeLa, HLF, and HEK293T cells were harvested by discarding used media, washing with PBS, adding 0.25% trypsin (Sigma) to just cover the vessel surface, placing cells at 37°C to detach, quenching trypsin with fresh media, and collecting the cell suspension. Semi-adherent MM1.S cells were harvested by both collecting media containing cells in suspension and detaching adherent cells using trypsin. Suspension CUTLL1 cells were harvested by collecting media. For passaging, cells were spun at 200×g for 3 minutes, resuspended in fresh medium, and seeded in new flasks. Cells were split 1:4 or 1:5 every 2–3 days.

Gill T., Wang H., Bandaru R., Lawlor M., Lu C., Nieman L.T., Tao J., Zhang Y., Anderson D.G., Ting D.T., Chen X., Bradner J.E, & Ott C.J. (2021). Selective targeting of MYC mRNA by stabilized antisense oligonucleotides. Oncogene, 40(47), 6527-6539.

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Personalized Predictive Cancer Models for MM

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We created ‘personalized’ predictive cancer models for MM cell line MM.1S (ATCC CRL-2974) and MM cell line U266B1 (ATCC TIB-196) as previously described^{10 (link),28 (link)–30 (link)} and were similar to the models listed in Supplementary Fig. S1. Cell line-specific genomic data was annotated into MM computational models. Cell associated biomarkers CD47, FASL, and PD-L1 and cell-free chemokines and cytokines IL6, IL-10 TGFB1, and VEGFA responses were predicted. The predicted responses from MM.1S and U266B1 were compared to each other and to the observed responses from MM.1S and U266B1 grown in culture.
Computational models of U266B1 and MM.1S were combined with DC to create MM.1S + DC and U266B1 + DC multi-cell computational models to predict the ability of these cells to inhibit DC marker expression. Connections integrated the common inputs and outputs representing paracrine and autocrine signaling between the cell types. Percent change in each output measure (with respect to the control network) was calculated after considering the output of MM cell lines as input into the DC single cell computational model. The magnitude and direction of these predictions was determined. DC biomarker responses for CD80, CD86, IL2, IFNG, and IL12B were predicted. Match rates were measured to assess whether U266B1 or MM.1S inhibited DC marker expression more in co-culture.

Fischer C.L., Bates A.M., Lanzel E.A., Guthmiller J.M., Johnson G.K., Singh N.K., Kumar A., Vidva R., Abbasi T., Vali S., Xie X.J., Zeng E, & Brogden K.A. (2019). Computational Models Accurately Predict Multi-Cell Biomarker Profiles in Inflammation and Cancer. Scientific Reports, 9, 10877.

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Cytotoxicity Assessment of Novel Compounds

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RPMI-8226, MM.1S (ATCC, Manassas, VA), or ALMC-2 (obtained from Dr. Diane Jelinek, Mayo Clinic, Rochester, MN) cells were plated (2.5 ×10⁴ cells/100 μL/well) in 96-well plates in the presence or absence of the novel compounds and incubated for 48 hrs. MTT solution (35 μL/well; 5 mg/mL PBS) was subsequently added and cells were incubated for an additional 4 hrs. MTT solubilizing solution (0.01 M HCl/10% SDS) was then added (100 μL/well). After overnight incubation at 37 °C, plates were analyzed on a microplate spectrophotometer at 540 nm. Compounds were tested in quadruplicate and three independent experiments were performed. The absorbance for control cells was defined as an MTT activity of 100%.

Foust B.J., Allen C., Holstein S.A, & Wiemer D.F. (2016). A New Motif for Inhibitors of Geranylgeranyl Diphosphate Synthase. Bioorganic & medicinal chemistry, 24(16), 3734-3741.

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Cultivation of Diverse Cell Lines

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The MM cell lines, MM1S, OPM2, OPM1, H929, OCIMY5, RPMI, U266, KMS1, HSB2, McCAR and ANBL6, and a breast cancer cell line MDA-MB-231 were obtained from ATCC (Manassas, VA). The T2 cell line, a human B and T cell hybrid expressing HLA-A2 molecules, was provided by Dr. J. Molldrem (University of Texas M. D. Anderson Cancer Center, Houston, TX). The cell lines were cultured in DMEM (for MM and T2 cells; Gibco-Life Technologies, Rockville, MD) or Leibovitz’s L-15 (for MDA-MB231; ATCC, Manassas, VA) media supplemented with 10% fetal calf serum (FCS; BioWhittaker, Walkersville, MD), 100 IU/ml penicillin and 100 µg/ml streptomycin (Gibco-Life Technologies).

Bae J., Samur M., Richardson P., Munshi N.C, & Anderson K.C. (2019). Selective Targeting of Multiple Myeloma by B cell Maturation Antigen (BCMA)-specific Central Memory CD8+ Cytotoxic T Lymphocytes: Immunotherapeutic Application in Vaccination and Adoptive Immunotherapy. Leukemia, 33(9), 2208-2226.

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Characterization of Multiple Myeloma Cell Lines

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Human MM cell lines MM.1S, U266, and NCI-H929 were obtained from ATCC. The human MM cell line MOLP-8 was purchased from DSMZ. Human MM cell lines KMS-11 and KMS-20 were purchased from the JCRB Cell Bank. Human MM cell line OPM-1 is a gift from Edward Thompson (University of Texas, Galveston, USA). The identities of MM.1S, U266, OPM-1, NCI-H929, and KMS-11 were validated by STR profiling (GenePrint®10 System, Promega). MOLP-8 cells expressing TurboGFP and luciferase (MOLP-8-TurboGFP-Luc) were generated by retrovirally transducing TurboGFP-IRES-luciferase bicistronic expression vector into MOLP-8 cells. Human embryonic kidney cell line 293T, human breast cancer cell line MCF7, and human colon cancer cell line Caco2 were otatined from ATCC. Cell lines were used within 3 months after thawing. Mycoplasma contamination was excluded using the MycoAlert Mycoplasma Detection Kit (Lonza). All MM cell lines were maintained in RPMI-1640 containing 100 U/ml penicillin and 100 μg/ml streptomycin, supplemented with 10% (v/v) FBS and 2 μM L-glutamine in 5% CO2 at 37 ^oC. 293T and MCF7 cells were maintained in DMEM supplemented with 10% (v/v) FBS. Caco2 cells were maintained in EMEM supplemented with 20% (v/v) FBS.

Ohguchi H., Park P.M., Wang T., Gryder B.E., Ogiya D., Kurata K., Zhang X., Li D., Pei C., Masuda T., Johansson C., Wimalasena V.K., Kim Y., Hino S., Usuki S., Kawano Y., Samur M.K., Tai Y.T., Munshi N.C., Matsuoka M., Ohtsuki S., Nakao M., Minami T., Lauberth S., Khan J., Oppermann U., Durbin A.D., Anderson K.C., Hideshima T, & Qi J. (2021). Lysine Demethylase 5A Is Required for MYC-Driven Transcription in Multiple Myeloma. Blood Cancer Discovery, 2(4), 370-387.

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Epigenetic Modulators in Hematological Cancers

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Human cancer cell lines K562, KMS11 and MM1S were purchased from ATCC. Antibodies were obtained from the following sources: EP300 (C-20) sc-585 from Santa Cruz, CREBBP (A-22) sc-369 from Santa Cruz, H3K27ac ab4729 from Abcam, MYC (N-262) sc-764 from Santa Cruz, GATA1 (N6) sc-265 from Santa Cruz, ACTB (Ac-15) A5441 from Sigma-Aldrich, PARP 9542 and Caspase3 9662 from Cell Signaling. CBP30 and I-CBP112 were purchased from Tocris Bioscience. C646 was purchased from Sigma-Aldrich. JQ1 was purchased from Selleck Chemicals. GNE-272 and CPI644 were synthesized in house. A-485 was obtained from the Structural Genomics Consortium.

Garcia-Carpizo V., Ruiz-Llorente S., Sarmentero J., Graña-Castro O., Pisano D.G, & Barrero M.J. (2018). CREBBP/EP300 bromodomains are critical to sustain the GATA1/MYC regulatory axis in proliferation. Epigenetics & Chromatin, 11, 30.

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