CBMN assessment of carotenoids in HepG2 cells

Description

This dataset contains micrographs of HepG2 cells exposed for 24h to the following carotenoids: 4.6 µM β-carotene, 40 µM trans-β-apo-8’-carotenal and 10 µM β-ionone 10 µM. Controls included cells grown only in culture medium, cells grown in solvent control (0.5% acetone) and cells grown in positive control 20 µM methyl methanesulfonate (MMS), a mutagenic compound. Cells were further incubated with cytochalalin B for a sufficient period of time to allow nuclear division while preventing cytokinesis.

Steps to reproduce

In 6-well plates, 5 × 10(5) HepG2 cells were seeded per well. After 24h, complete DMEM medium (10% FBS) was replaced with medium containing one of the following: - 40 µM trans-β-apo-8’-carotenal; - 4.6 µM β-carotene; - 10 µM β-ionone; - 0.5 % acetone (v/v) - 20 µM methyl methanesulfonate - control (complete DMEM only) After 24h, the medium was replaced with medium containing 3 µg/mL cytochalasin B, to prevent cytokinesis. After 30h, cells were washed with PBS and detached with 0.25% trypsin - 1 mM EDTQA solution. Cells in suspension, 5 × 10(3), were centrifuged and spotted on glass slides. After methanol fixation (10 min) and drying, cells were stained sequentially with Eosin Y dye and Azur B from Hemacolor staining kit, according to the kit's instructions.

Institutions

Categories

Funding

Licence