Quantitative Live-Cell Analysis Using Automated Long-Term Imaging
Poster Sep 13, 2017
Joe Clayton and Peter Banks
Characterizing cell proliferation is a crucial aspect of biological research and therapeutic drug development. Most current cell proliferation assays rely on indirect biochemical metrics that are limited by artifacts or imaging-based endpoint measures.
Here we describe a continuous live-cell assay for determining cell proliferation profiles using the BioSpa Automated Live Cell Imaging System, consisting of BioSpa 8 and Cytation 5. This fully automated method enables quantitative and phenotypic long-term analysis of cell growth using non-invasive measures of confluence or direct cell count. To demonstrate the abilities of this system to conduct robust and reproducible kinetic proliferation assays, NIH3T3, HeLa, and HCT116 cell growth was followed for five days. All three cell types exhibited robust logarithmic growth up to full confluence with doubling times consistent with literature values. Additionally, to demonstrate the ability of this system to screen pharmacological agents, cell proliferation profiles for cells cultured with eight concentrations of two literature-standard inhibitory compounds were generated. Calculated IC50 values were used to measure drug response for each compound and cell type.
Multiplexing cell-based assays is possible using 3D culture models that are larger and more complex than monolayers
Real-time detection methods to measure live or dead cells provide much flexibility for multiplexing
All multiplexed assay combinations should be verified using appropriate controls for each 3D cell culture model.
Basic fibroblast growth factor (bFGF) is widely used in vitro for the maintenance and stimulation of a variety of cells. However, use of native bFGF in cell biology is limited by the fact that bFGF rapidly degrades at physiological temperatures. We have addressed this problem with an engineered form of bFGF, named Heat Stable bFGF (HS bFGF), which is stable at 37 degrees Celsius.READ MORE