Peripheral blood mononuclear cells (PBMC) were obtained by density-gradient centrifugation and red blood cell lysis from peripheral blood of healthy donors in agreement with institutional consent and collection guidelines (Institute for Transfusion Medicine and Haemotherapy, University Hospital Wurzburg and Wurzburg University Ethic Committee, No. 141/17-sc). The human cell lines THP-1 (acute myeloid leukemia, ATCC, No. TIB-202 and DSMZ, No. ACC-16), HT1080 (sarcoma, ATCC, No. CCL-121), U266 (multiple myeloma, ATCC, No. TIB-196), Jurkat (T cell leukemia, DSMZ, No. ACC-282), KMS-12-BM (multiple myeloma, DSMZ, No. ACC-551), Raji (Burkitt lymphoma, ATCC, No. CCL-86), and MDA-MB-231 (ATCC, No. HTB-26) were grown in advanced RPMI-1640 or advanced DMEM supplemented with 200 μM l -glutamine, 10% FBS, penicillin (200 U/mL), and streptomycin (200 μg/mL) (Thermo Fisher Scientific, MA, USA). We routinely tested cell lines for absence of mycoplasma infection. Authentication of the cells was performed by the provider.
Kms12bm
Manufactured by American Type Culture Collection
Sourced in United States
The KMS12BM is a laboratory equipment designed for the cultivation and maintenance of cells. It provides a controlled environment for the growth and development of cell cultures, ensuring optimal conditions for their survival and proliferation. The core function of the KMS12BM is to maintain the appropriate temperature, humidity, and gas composition required for the specific cell lines being cultivated.
Automatically generated - may contain errors
Lab products found in correlation
Mm 1s, by American Type Culture Collection (4 mentions)
Rpmi8226, by American Type Culture Collection (3 mentions)
Nci h929, by American Type Culture Collection (3 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Skmm1, by American Type Culture Collection (2 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
Advanced dmem, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Advanced rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
Ina 6, by American Type Culture Collection (1 mentions)
Myco lumi luminescent mycoplasma detection kit, by Beyotime (1 mentions)
Snu216, by Cobioer Biosciences (1 mentions)
Kms 12 pe, by American Type Culture Collection (1 mentions)
Nci n87, by Cobioer Biosciences (1 mentions)
Hgc 27, by Cobioer Biosciences (1 mentions)
Snu 719, by Cobioer Biosciences (1 mentions)
Kms 34, by American Type Culture Collection (1 mentions)
7 protocols using kms12bm
1
Isolation and Culture of Human Immune Cells
2
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.
3
Culturing Human Cancer Cell Lines
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Human GC cell lines, including 23132/87, SNU216, NCI‐N87, MKN1, AGS, HGC27, and SNU719, were purchased from Cobioer (Nanjing). Human multiple myeloma cell lines, including MM1S, KMS12PE and KMS12BM, were purchased from ATCC and DSMZ. All cancer cell lines were cultured in RPMI‐1640 medium (Gibco, CA, USA) supplement with 10% fetal bovine serum (FBS) (Gibco, CA, USA) at 37°C with 5% CO2. The human embryonic kidney cell line HEK293T was cultured in DMEM supplemented with 10% FBS at 37°C with 10% CO2. Cells were tested regularly using the Myco‐Lumi Luminescent Mycoplasma Detection Kit (Beyotime, C0297M)and were consistently negative for mycoplasma.
4
Cell Line Authentication and Culture
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Cell lines and cell culture KMS11, KMS18, KMS34, H929, OPM2, KMS12BM, U266, and RPMI8226 cells were purchased from ATCC and cultured in RPMI1640 medium supplemented with 10% FBS, 100 U/mL penicillin, and 100 mg/mL streptomycin for 1-2 weeks before experimental use. TKO cells are derived from t(4;14)þ KMS11 cells, where the translocated IGH-NSD2 allele has been inactivated by homologous recombination to retain one copy of the wild-type NSD2 allele (nontranslocated allele). HEK293T cells were cultured in DMEM supplemented with 10% FBS, 100 U/mL penicillin, and 100 mg/mL streptomycin. All of the listed cells above were cultured at 37 C and 5% CO 2 . Cell lines were periodically tested for Mycoplasma (Lonza) and authenticated (Centre for Translational Research and Diagnostics, National University of Singapore, Singapore).
5
Manipulating miR-125b and MALAT1 in MM Cells
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Peripheral blood mononuclear cells including PBMC1, PBMC2, PBMC3, and PBMC4, were donated by normal healthy individuals. Other eight MM cell lines (NCI‐H929, U266, SKMM1, MMIS, KMS‐12‐BM, MM1R, PRMI‐8226, and INA‐6) were purchased from American Type Culture Collection. All cell lines were maintained in RPMI‐1640 (Life Technologies, Carlsbad, CA) supplemented with 10% fetal bovine serum (Lonza Group Ltd., Switzerland) and 1% penicillin/streptomycin (Life Technologies) with 5% humidified CO2 at 37°C. The miR‐125b overexpression lentiviral vector (Lenti‐miR‐125b), miR‐125b down‐regulation lentiviral vector (Lenti‐Sh‐miR‐125b), MALAT1 Si‐RNA (Si‐MALAT1), and MALAT1 overexpression vector (pcDNA ‐MALAT1) were purchased from Qiagen (Hilden, Germany). After cultured in six‐well plates supplemented with antibiotic‐free medium, NCI‐H929 and PRMI‐8226 cells were transfected with lentiviral supernatant to obtain stable‐transfected cell lines and chosen for the following procedures.
6
High-Throughput Screening of Multiple Myeloma Cell Lines
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KMS-12-BM, NCI-H929, U-266, SK-MM-1, OPM-2, and MM.1S cell lines were purchased from the American Type Culture Collection (ATCC, Wesel, Germany). All cell lines were cultured in RPMI-1640 medium, supplemented with 10–20% heat-inactivated fetal bovine serum (FBS), 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/ml streptomycin (Gibco, Thermo Fisher Scientific, Waltham, MA, United States) at 37°C in a humidified incubator with 5% CO2. The compounds were pre-plated onto 384-well plates in five concentrations in a 10,000-fold concentration range. The cells were seeded in 25 μl volume of medium at the density of 5000 cells/well. After 72 h, cell viability was measured using the CTG assay. The data were normalized to negative control (dimethyl sulfoxide vehicle only) and the positive controls wells (100 μmol/l benzethonium chloride).
7
Culturing Multiple Myeloma Cell Lines
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We used four well‐known MM cell lines with various molecular subtypes: RPMI‐8226, KMS‐12‐BM, KMS‐11, and MM.1 S. These cell lines were purchased from the American Type Culture Collection (ATCC). SACHI and SK‐MM‐1 were kindly provided by Dr. Ichiro Hanamura (Aichi Medical University, Aichi, Japan). These cell lines were cultured in RPMI 1640 medium (Thermo Fisher Scientific) supplemented with 10% inactivated fetal calf serum (FCS). 293FT cells were cultured in DMEM containing 10% inactivated fetal calf serum. A multi‐gas incubator MCO‐5 M‐PJ (PHC, Tokyo, Japan) was used for hypoxic culture (1% O2).
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