The acid phosphatase (ACP) assay was used to measure cell viability as described previously (12 (link)). Briefly, 5000 cells/well in the 96-well plate were seeded and incubated with different concentrations of glucose or treated with Mdivi-1 (10 μM) for 3-days. Alternatively, clonogenic cell survival was performed 48-hr after nutrient deprivation. Cells were trypsinized, combined with suspended cells, and total 100 cells were reseeded in 6-well plate for 2-weeks as described previously (7 (link)). Apoptosis was analyzed by Annexin V/Propidium iodide (PI) staining (BD Pharmingen) as described previously (12 (link)).
Protocol detail
Assessing Cell Viability and Survival
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Corresponding Organization : City of Hope
Other organizations : Chi Mei Medical Center, Southern Taiwan University of Science and Technology, California Institute of Technology, University of California, Davis, National Health Research Institutes
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Variable analysis
independent variables
- Different concentrations of glucose
- Mdivi-1 (10 μM)
- Cell viability (measured by acid phosphatase (ACP) assay)
- Clonogenic cell survival (after 48-hr nutrient deprivation)
- Apoptosis (analyzed by Annexin V/Propidium iodide (PI) staining)
- Cell seeding density (5000 cells/well in 96-well plate)
- Incubation time (3-days)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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