A Theoretical and Experimental Study to Optimize Cell Differentiation in a Novel Intestinal Chip

Area of Science:

• Biomedical Engineering
• Cell Biology
• Microfluidics

Background:

• Trans-well cultures, a common in vitro gut model, lack physiological shear stress crucial for Caco-2 cell differentiation into enterocytes.
• Organ-on-chip models offer a promising alternative by providing necessary shear stress for cell culture.

Purpose of the Study:

• To develop a novel 3D-printed microfluidic device that applies physiologically-relevant shear stress to Caco-2 cells.
• To compare cell differentiation and intestinal barrier function under pump-driven flow versus gravity-driven flow.

Main Methods:

• A biocompatible 3D-printed microfluidic device was designed and fabricated.
• Caco-2 cells were cultured under physiologically-relevant unidirectional shear stress and compared to gravity-driven flow conditions.
• Numerical studies calculated flow rates for desired shear stress, and experimental tests verified effects on cell differentiation.

Main Results:

• The pump-driven flow system (PDFS) significantly enhanced Caco-2 cell differentiation compared to the gravity-driven flow system (GDFS).
• Computational modeling confirmed the device's ability to provide homogeneous shear stress, potentially improving differentiation.
• The microfluidic device promoted tighter monolayers and enhanced functional properties of differentiated Caco-2 cells.

Conclusions:

• The novel microfluidic device effectively mimics physiological shear stress, promoting Caco-2 cell differentiation and intestinal barrier function.
• This technology presents a promising animal-free platform for preclinical in vitro drug testing.