Cell stimulus and lysis in a microfluidic device with segmented gas-liquid flow

Area of Science:

• Cell biology
• Biotechnology
• Microfluidics

Background:

• Understanding rapid cell signaling requires precise control over stimulus delivery and response measurement.
• Existing methods may lack the speed and resolution needed for transient cellular network dynamics.

Purpose of the Study:

• To develop and validate a microfluidic device for rapid mammalian cell stimulation and lysis.
• To investigate the impact of segmented gas-liquid flow on cell responses and signaling pathways.
• To resolve fast transient responses in cell signaling networks with high fidelity.

Main Methods:

• Utilized a microfluidic device employing segmented gas-liquid flow for enhanced mixing.
• Integrated thermoelectric elements for precise temperature control during cell stimulation and lysis.
• Assessed cellular responses in three cell types, including morphological and stress-activated kinase analyses.
• Stimulated Jurkat E6-1 cells with anti-CD3 to measure Extracellular signal-regulated Kinase (ERK) and c-Jun N-terminal Kinase (JNK) activation over time.

Main Results:

• The microfluidic device demonstrated rapid stimulus and lysis capabilities.
• Segmented flow did not induce morphological changes or activate stress-sensitive mitogen-activated protein kinases (MAPKs) in tested cell types.
• On-chip stimulation of Jurkat cells resulted in ERK and JNK pathway activation profiles identical to conventional stimulation methods.
• Validated the device's ability to capture fast transient responses in cell signaling.

Conclusions:

• The developed microfluidic device accurately resolves fast transient responses in cell signaling networks.
• The device provides a robust platform for studying cellular dynamics without compromising cell integrity or function.
• This technology offers a valuable tool for advancing research in cell signaling and network dynamics.