Simultaneous Plate-Reader Characterization of Promoter Activity and Cell Growth in Engineered Mammalian Cells

Area of Science:

• Biotechnology
• Cell Biology
• Assay Development

Background:

• Automated high-throughput methods are crucial for mammalian cell line characterization and genetic engineering.
• Current methods for tracking cell growth can be labor-intensive and invasive.
• There is a need for efficient, noninvasive assays to monitor cell proliferation.

Purpose of the Study:

• To develop and validate a high-throughput, noninvasive assay for tracking mammalian cell growth.
• To demonstrate the utility of phenol red absorbance changes for quantifying cell proliferation.
• To establish a computational pipeline for predicting cell numbers from plate reader data.

Main Methods:

• Utilized a plate reader to measure changes in absorbance of the pH indicator phenol red.
• Developed a computational pipeline to convert absorbance profiles into a cell growth index.
• Calibrated measurements to enable prediction of cell numbers from absorbance data alone.

Main Results:

• Phenol red's basic and acidic absorbance profiles correlate with cell count.
• The assay accurately predicts cell numbers using plate reader measurements.
• The method is effective for both suspension and adherent cell lines under various conditions.

Conclusions:

• The developed assay provides a high-throughput, noninvasive method for mammalian cell growth tracking.
• This approach facilitates cell line characterization, genetic engineering, and drug screening.
• The assay supports the identification of desired promoters in engineered cell lines.