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.
We found a distinct subpopulation of Tregs within BMSCs. Tregs and BMSCs in co-culture conferred neuroprotection that varied in a dose-dependent manner. Tregs minimized stem cell production of IL-6, a pro-inflammatory cytokine, and inhibited BMSC secretion of FGF-beta, a cytokine related to BMSC proliferation and differentiation. The ratio of Tregs found natively in BMSCs is optimally adapted to provide the maximum neuroprotective benefit of stem cell treatment after ischemic stroke.READ MORE
We utilized paired synthetic crRNAs coupled with our synthetic tracrRNA in cells transduced with lentiviral Cas9 to perform a functional knockout on hsa-miR-221. This three-part system (crRNA, tracrRNA and Cas9) has demonstrated efficient gene editing when used with only one guide RNA, but the goal was to use two crRNAs to remove the entire stem-loop.READ MORE
2nd International Conference on Pharmaceutical Research & Innovations in Pharma Industry
May 30 - May 31, 2019