Cell viability test for the membrane-specific drug delivery
This protocol is extracted from research article:
Transport of a graphene nanosheet sandwiched inside cell membranes
Sci Adv, Jun 7, 2019; DOI: 10.1126/sciadv.aaw3192

The cytotoxicity of the loading drugs (VTB and REGO) was tested by a cell counting kit (CCK8). Typically, 5000 cancer cells (4T-1 and MCF-7) were cultured in each well (100 μl) of a 96-well plate and allowed to adhere overnight. Cells were then treated with 10-μl serial dilutions of drug or carrier-loaded drug. We also prepared wells for background absorbance measurement that contains all empty carriers except cells. After 24 hours of cell-drug incubation, 10 μl of CCK8 was added to the cells and incubated for 1 hour at 37°C. The absorbance of a water-soluble formazan product was determined on an Infinite M200 microplate spectrophotometer (Tecan) at 450 nm, which was normalized to compare with the untreated cell control. For further confirmation, cells were seeded on a petri dish, and the cell viability was detected by a LIVE/DEAD kit. On the basis of good esterase activity of live cells and impaired membrane of dead cells, live (stained by green calcein) and dead (stained by red EthD-1) cells were imaged by a confocal imaging system (Leica TCS SP5).

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