Label-Free Single Microparticles and Cell Aggregates Sorting in Continuous Cell-Based Manufacturing

Area of Science:

• Biotechnology
• Bioengineering
• Cell Biology

Background:

• Continuous bioprocessing is shifting biomanufacturing paradigms.
• Cell-based manufacturing faces challenges in real-time monitoring and harvesting of adherent and suspension cultures.
• Existing methods lack efficient tools for in-line analysis of biomass, cell viability, and differentiation.

Purpose of the Study:

• To develop a novel label-free microfluidic platform for high-throughput impedance bioanalysis.
• To integrate real-time particle sorting based on multi-frequency impedance signatures.
• To enable automated analysis of cell quality attributes for continuous cell-based manufacturing.

Main Methods:

• A microfluidic platform utilizing impedance bioanalysis for high-throughput (≈50 particles/sec) analysis.
• Integration of a real-time piezo-actuated particle sorter.
• Application of multi-frequency impedance signatures for biomass profiling, cell viability assessment, and stem cell differentiation analysis.

Main Results:

• Successfully profiled biomass of Cytodex-3 microcarriers with adipose-derived mesenchymal stem cells (ADSCs) for sorting.
• Demonstrated impedance-based isolation of microcarriers with osteogenically differentiated ADSCs, validated by increased calcium content.
• Performed impedance profiling of heterogeneous ADSCs-encapsulated hydrogel microparticles and 3D ADSC aggregates to sort for high biomass and viability.

Conclusions:

• The scalable microfluidic platform enables in-line sample processing directly from bioreactors.
• Automated analysis of cell quality attributes maximizes cell yield and improves control in continuous cell-based manufacturing.
• This technology addresses critical needs in bioprocessing for enhanced cell quality and efficient production.