Use of cryopreserved cells for enabling greater flexibility in compound profiling

Area of Science:

• Biochemistry
• Pharmacology
• Cell Biology

Background:

• Compound profiling assays commonly measure intracellular calcium release using the Fluorometric Imaging Plate Reader (FLIPR).
• Traditional FLIPR assays rely on cells in continuous culture, limiting assay execution to weekdays and requiring specialized automation for weekend plating.
• Iterative chemistry campaigns necessitate efficient and flexible compound profiling methods.

Purpose of the Study:

• To develop a method for generating high-quality compound profiling data using cryopreserved cells.
• To demonstrate that cryopreserved cells can yield equivalent data to continuously cultured cells.
• To enable compound profiling assays throughout the entire week without continuous cell culture.

Main Methods:

• Utilized cryopreserved cells for intracellular calcium release assays.
• Compared data quality from cryopreserved cells plated at various densities and time points (24-72 h post-plating).
• Assessed the feasibility of generating structure-activity relationship (SAR) data using the cryopreserved cell method.

Main Results:

• High-quality compound profiling data was successfully generated from cryopreserved cells.
• Equivalent data quality was achieved between 24 and 72 hours post-plating across different cell densities.
• The method demonstrated robustness for deriving structure-activity relationship (SAR) data.

Conclusions:

• Cryopreserved cells offer a viable alternative to continuous culture for FLIPR-based compound profiling.
• This approach enhances assay flexibility, allowing data generation throughout the week.
• The method reduces reliance on complex cell culture automation and continuous culturing, optimizing drug discovery processes.