Can you create a protocol based off the information given

Can you create a protocol based off the information given
NB-5 NB-6 ● Amsa 20 40 Compound Concentration (uM) 60 80 MIT Assay The cytotoxicity of the compounds was tested against nine human tumor cell lines: HL-60 (acute promyelocytic leukemia). HL60/MX1 (mitoxantrone-resistant acute promyelocytic leuke- mia). CCRF-CEM (acute lymphoblastic leukemia), NG97 (glyo- blastoma) T47-D (mammary ductal adenocarcinoma), Raji Burkitts lymphoma) and Jurkat (T-cell leukemia). All cell lines were obtained from Rio de Janeiro Tissue Cell Bank except NG97 cells that were kindly provided by University of São Paulo. Cells were cultured in RPMI-1640 medium, supplemented with 10% fetal calf serum, 2 mM glutamine, 100mg/mL streptomycin and 100 U/mL penicillin at 37 C with 5% CO2-For experiments, cells were plated in 96-well plates (10 cells/well. After 24h, the compounds (1 to 75 μM) dissolved in DMSO were added to each well and incubated for 3 days (72h). Control groups received the same amount of DMSO (0.1%). Amsacrine was used as positive control. Growth of tumor cells was quantified by the ability of living cells to reduce the yellow dye 3-(4,5-dimethyl-2-thiazolyl)- 2.5-diphenyl-2H-tetrazolium bromide (MIT) to a blue formazan product. At the end of 72 h incubation, 20 μLof 0.5 mg/mL of MTT were added in each well, and cells were incubated again at 37 C with 5% CO-Three hours later, the formazan product of MTT reduction was dissolved in 20% SDS, and absorbance was measured using a multi-plate reader (EL808, Biotek, USA). Drug effect was quantified as the percentage of control absorbance of reduced dye at 570 nm.

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