An integrated microfluidic bubble pocket for long-term perfused three-dimensional intestine-on-a-chip model

Area of Science:

• Biotechnology
• Developmental Biology
• Microfluidics

Background:

• Perfused three-dimensional (3D) cultures support long-term organoid growth and function.
• Air bubbles disrupt long-term, on-chip perfused 3D organoid cultures, limiting their utility.
• Investigating organoid development, growth, and function requires stable culture conditions.

Purpose of the Study:

• To develop an imaging platform that prevents air bubble formation in long-term perfused 3D organoid cultures.
• To enable stable, long-term monitoring of gastrointestinal (GI) organoid development and function.
• To overcome the limitation of disruptive air bubbles in microfluidic organoid culture systems.

Main Methods:

• Utilized 3D printing to create polymer molds for polydimethylsiloxane (PDMS) culture chambers.
• Integrated an innovative microfluidic bubble pocket into the PDMS culture chambers.
• Developed a platform for long-term perfused 3D culture of GI organoids with bubble trapping and diffusion mechanisms.

Main Results:

• The developed platform successfully trapped and diffused unintended air bubbles in the integrated PDMS pocket.
• The platform effectively prevented bubble formation in human and mouse GI organoid cultures during long-term perfusion.
• Demonstrated robust, long-term perfused time-course imaging of GI organoids without bubble disruption.

Conclusions:

• The novel imaging platform with an integrated bubble pocket effectively circumvents air bubble issues in long-term perfused GI organoid cultures.
• This robust platform is ideally suited for studies requiring long-term perfusion monitoring of organoid growth, morphogenesis, and function.
• The technology facilitates advanced research in organoid development and disease modeling.