On-chip incubation system for long-term microfluidic cell culture

Area of Science:

• Biotechnology
• Microfluidics
• Cell Biology

Background:

• Traditional cell culture often requires complex external equipment for controlled environments.
• Developing integrated systems for cell culture can streamline experiments and improve reproducibility.

Purpose of the Study:

• To demonstrate a microfluidic cell culture chip with on-chip CO2 incubation.
• To showcase a system that negates the need for external gas supply or incubation chambers.
• To validate the chip's performance for long-term cell culture.

Main Methods:

• Fabrication of a poly(dimethylsiloxane) (PDMS) microfluidic chip with nested reservoirs and a water-jacket.
• Utilizing a sodium bicarbonate and sodium carbonate solution as the water-jacket for CO2 control.
• Employing Braille pin-driven pumping for fluid manipulation.
• Culturing CV-1 epithelial cells on-chip and monitoring under an inverted microscope.

Main Results:

• The chip successfully maintained stable pCO2 levels (5.0% ± 0.2%) and osmolality (< 3 mmol/kg) for over 24 hours.
• Long-term microfluidic cell culture of CV-1 epithelial cells was achieved for at least 12 days.
• The integrated system demonstrated effective on-chip CO2 incubation without external gas supply.

Conclusions:

• The developed microfluidic chip offers a self-contained solution for cell culture with controlled CO2 incubation.
• This technology has the potential to simplify cell-based assays and advance microfluidic cell culture applications.
• The system provides a stable microenvironment for extended cell culture studies.