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Microfluidic Buffer Exchange for Interference-free Micro/Nanoparticle Cell Engineering

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

Lingyan Gong¹, Linwei He¹, Nan Lu¹

¹School of Mechanical and Aerospace Engineering, Nanyang Technological University, Singapore, 639798, Singapore.

Advanced Healthcare Materials

|March 11, 2024

Summary

This study introduces a novel microfluidic platform for real-time monitoring and sorting of cells in continuous bioprocessing. The technology enhances cell quality and yield by enabling precise analysis of biomass and viability.

Keywords:

cell aggregates cell‐based manufacturing hydrogel microparticles impedance cytometry microcarriers microfluidics sorting

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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.