Abstract
Combinations of living cell-based biosensors and microdevices are attractive tools for real-time monitoring of gene expression profiling in a small population of cells involving small amount of analytes. However, due to the heterogeneous responsiveness of cells, cell-based biosensors have poor reproducibility and a low signal-to-noise (S/N) ratio. Previously, we constructed a cell, a GFP reporter cell line containing an engineered Heart Shock Protein 70B' promoter generated by stably transfecting mouse NIH/3T3 cells. In this study, we manipulated the cell density to overcome the lower signal and poor reproducibility using the sensor cells. We found that a cell density of 2 x 10 5 cells/cm 2 provides good responsiveness of sensor cells that appears to be related to the G0/G1 phase of cell cycle. However, higher cell densities had a negative effect for on sensor performance. We also designed microdomains to regulate cell density. The GFP-positive rate of cells grown on domains at 2 x 10 5 cells/cm 2 density was approximately 1.5 times higher than that of control cells. Our results suggest that cell density is an important factor for the design of cell-based biosensors with microdevices.
Highlights
During initial investigations of an undefined substance that may be diffused by chemical or biological terrorism, identification of the substance is important
Approximately 80% of cells respond when synchronized in the G0/G1 phase using serum starvation treatment. These results suggest that cell cycle synchronization is an important tool to enhance the responsiveness of cell-based biosensors and facilitate good reproducibility
The results indicate that cell density is an important factor for cellular responsiveness and that consideration must be given to cell density when designing cell-based biosensor microdevices
Summary
During initial investigations of an undefined substance that may be diffused by chemical or biological terrorism, identification of the substance is important. In this regard, living cell-based biosensors have been developed to determine the effects of extracellular chemical or/and physical stimuli through cellular function. These results suggest that cell cycle synchronization is an important tool to enhance the responsiveness of cell-based biosensors and facilitate good reproducibility.
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