Cytotoxicity Assay for Mammalian Cell Lines

Example 43

Mammalian cells (HepG2, A549, and HEK293) were obtained from the ATCC and cultured routinely in DMEM with 10% FBS and 1% (v/v) antibiotic-antimycotic. For cytotoxicity assays, 1×10⁶cells were seeded into 384-well plates 1 day before compound treatment. Cells were treated with ascending doses of compound for 72 h, and viability was measured using Cell-Titer Glo (Promega). All cell lines were tested for Mycoplasma contamination using Universal mycoplasma Detection Kit (ATCC).

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US11866441B2. Compounds and methods for the treatment of parasitic diseases (2024-01-09). THE BROAD INSTITUTE, INC. [US], PRESIDENT AND FELLOWS OF HARVARD COLLEGE [US]. Inventors: Eamon Comer [US], Nobutaka Kato [US], Marshall Morningstar [US], Bruno Melillo [US].

Patent 2024

Cell-Titer Glo Universal mycoplasma Detection Kit

Antibiotic Assays Cell Cell lines Cytotoxicity Mammalian Mycoplasma Promega

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Variable analysis

independent variables

Ascending doses of compound

dependent variables

Cell viability

control variables

Cell culture media (DMEM with 10% FBS and 1% antibiotic-antimycotic)
Cell seeding density (1×10^6 cells per well)
Incubation time (72 hours)
Mycoplasma contamination testing

positive controls

None specified

negative controls

None specified

Annotations

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