Simultaneous cell capture and induction of apoptosis using an anti-CD95 affinity microdevice

Area of Science:

• Biotechnology
• Cell Biology
• Microfluidics

Background:

• Apoptosis, or programmed cell death, is crucial for development and disease.
• Current methods for studying apoptosis can be limited in real-time observation and cell viability.
• Jurkat cells are a human T-cell line commonly used in apoptosis research.

Purpose of the Study:

• To design and demonstrate a microfluidic device for simultaneous cell capture and apoptosis induction.
• To investigate the use of anti-CD95 antibodies for both cell capture and apoptosis signaling.
• To enable real-time monitoring of apoptosis dynamics in Jurkat cells.

Main Methods:

• Fabrication of a poly(dimethylsiloxane) microfluidic channel sealed to glass.
• Coating the channel with anti-CD95 antibodies for affinity-based cell capture.
• Flowing medium through the channel to maintain cell viability during apoptosis induction.
• Staining with Annexin V-PE and 7-AAD for apoptosis detection.

Main Results:

• The microfluidic device successfully captured Jurkat cells via anti-CD95 binding to the Fas receptor.
• 43.5% of captured cells exhibited apoptosis after 3 hours, compared to 7.9% in conventional flow cytometry methods.
• The device maintained cell viability, allowing for long-term observation of apoptosis.

Conclusions:

• The developed microfluidic device offers a unique platform for simultaneous cell capture and apoptosis induction.
• This system provides a more efficient and observable method for studying apoptosis compared to traditional techniques.
• The device holds potential for future studies on the temporal dynamics of apoptosis and drug screening.