Automated image processing as an analytical tool in cell cryopreservation for bioprocess development.

Area of Science:

• Biotechnology and Cell Therapy
• Bioprocess Engineering
• Cryobiology

Background:

• Continuous cell availability is critical for biopharmaceutical and cell therapy industries.
• Long-term cell stability relies on optimized cryopreservation strategies, including controlled freezing, ultra-low temperature storage, and rapid thawing.
• Current empirical methods for cryopreservation strategy development are suboptimal; a knowledge-based approach is highly desirable.

Purpose of the Study:

• To develop and validate a video-based analytical tool for characterizing freezing and thawing behavior during cryopreservation.
• To enable a more knowledge-based approach to cryopreservation process development.
• To assess the tool's flexibility and practicality using a model cell line (INS-1E) and varying working volumes.

Main Methods:

• Development of a video-based tool to monitor freezing and thawing dynamics.
• Validation using a model cryopreservation process with the INS-1E cell line.
• Evaluation of performance across two working volumes (1.0 mL and 2.0 mL), assessing freezing-thawing rates, cell recovery, and biomass increase.

Main Results:

• The video-based tool successfully identified a delay in freezing and thawing for the larger working volume (2.0 mL), demonstrating its flexibility.
• A significant decrease in cell recovery was observed when increasing the working volume from 1.0 mL (0.94 ± 0.14%) to 2.0 mL (0.61 ± 0.05%), highlighting the tool's practicality.
• The tool accurately characterized the impact of working volume on cryopreservation process performance.

Conclusions:

• The developed video-based tool is a powerful asset for cell-specific optimization of cryopreservation protocols.
• It facilitates faster and more systematic cryopreservation process development by providing detailed insights into freezing and thawing behavior.
• This approach moves beyond empirical methods, enabling a more directed and knowledge-based strategy for creating stable cell banks.