To generate heat-induced clones, cells were first incubated at 42°C for 4 h over three consecutive days and then single-cell sorted (FACSAria II; BD Biosciences) based on cell size into individual wells in 96-well plates with 50% conditioned media. After ∼2–3 wk of incubation, colonies were harvested and their ploidy was analyzed by FACS and metaphase spreads. For each drug treatment, two biological replicates with two technical replicates per clone was conducted. Four hundred cells/well were plated a day before they were subjected to the indicated drug treatments for 3 h. After 1 wk of incubation under standard conditions, cells were ethanol-fixed and stained using 0.05% crystal violet (C3886; Sigma-Aldrich). Counting was performed manually. Drugs: bleomycin (ab142977; Abcam), cisplatin (ab141398; Abcam), daunorubicin (D0125000; Sigma-Aldrich), doxorubicin (D1515; Sigma-Aldrich), oxaliplatin (ab141054; Abcam), STLC (164739; Sigma-Aldrich), paclitaxel (T7402; Sigma-Aldrich), vinblastine (V1277; Sigma-Aldrich). All drugs were dissolved in dimethyl sulfoxide (DMSO) (D2650; Sigma-Aldrich) except cisplatin, which was dissolved in saline.
D0125000
Manufactured by Merck Group
Sourced in France
D0125000 is a laboratory equipment product manufactured by Merck Group. It serves as a general-purpose device for conducting various laboratory experiments and analyses. The core function of this product is to provide a controlled and standardized environment for scientific research and testing purposes. Detailed technical specifications and intended use cases are not available for this specific product.
Automatically generated - may contain errors
Lab products found in correlation
Daunorubicin, by Merck Group (1 mentions)
Vinblastine, by Merck Group (1 mentions)
Ab142977, by Abcam (1 mentions)
Doxorubicin, by Merck Group (1 mentions)
C3886, by Merck Group (1 mentions)
T7402, by Merck Group (1 mentions)
D1515, by Merck Group (1 mentions)
D2650, by Merck Group (1 mentions)
Facsaria 2, by BD (1 mentions)
Ara c, by Merck Group (1 mentions)
Hl 60, by Leibniz Institute DSMZ (1 mentions)
Molm 13, by Leibniz Institute DSMZ (1 mentions)
Fetal bovine serum (fbs), by PAN Biotech (1 mentions)
Pen strep, by PAN Biotech (1 mentions)
2 protocols using d0125000
1
Heat-Induced Clones Generation and Drug Sensitivity
2
Establishing AML Cell Line Resistance
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Acute Myeloid Leukemia (AML) cell lines HL-60, NB-4, MOLM13, and KG-1 were acquired from the DSMZ-German Collection of Microorganisms and Cell Cultures GmbH. Cells were maintained and routinely subcultured in culture flasks at a cell density between 1 × 105 and 1 × 106 cells/mL. Cells were grown in a complete medium consisting of RPMI (PanBiotech, Aidenbach, Germany) supplemented with 10% Fetal Bovine Serum (FBS, PanBiotech) and 1% penicillin/streptomycin (PenStrep, PanBiotech). Cell cultures were maintained in a humidified incubator at 37 °C and a 5% CO2 atmosphere. To induce resistance, cells were exposed to increasing concentrations of Cytarabine (Ara-C, C3350000, Sigma, Strasbourg, France) and Daunorubicin (DNR, D0125000, Sigma) for 3–6 months. To obtain enough biomass to perform the assays, cells were grown to high densities, but not exceeding 1 × 106 viable cells/mL and with cell viability up to 90%. Viable cell estimation was performed by adding 1:1 cell suspension to 0.2% trypan blue (sc-216028, Santa Cruz Biotechnology, Dallas, TX, USA), and cells were counted in a Neubauer chamber.
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