Microfluidic-based gelatin methacrylate microgel as a scaffold to create reverse-polarity HT29 spheroids

Area of Science:

• Biomedical Engineering
• Cell Biology
• Gastroenterology

Background:

• In vitro models of the human gut are crucial for predicting drug absorption and toxicity.
• Understanding host-pathogen interactions in infectious diseases requires accurate gut models.
• Existing models often lack the complex microarchitecture of the intestinal villi.

Purpose of the Study:

• To develop a novel in vitro human gut model using microgels.
• To investigate cell behavior and structure on a biomimetic intestinal surface.
• To assess the potential of this model for drug absorption, toxicity, and infectious disease studies.

Main Methods:

• Gelatin methacrylate (GelMa) microgels were fabricated using flow-focusing droplet microfluidics.
• Microgels of approximately 80 μm size were produced with controlled stiffness (0.01 kPa).
• HT29 cells were cultured on microgels to form reverse-polarity spheroids, termed the 'microgut'.

Main Results:

• HT29 cells cultured on microgels maintained high viability (95%).
• Cells exhibited polarization in response to microgel curvature, with nuclear orientation and F-actin expression.
• HT29 cells on GelMa7 microgels showed enhanced tight junction expression compared to other concentrations.

Conclusions:

• The developed 'microgut' model effectively replicates intestinal villi microarchitecture.
• This model demonstrates cellular polarization and enhanced tight junction formation.
• The microgut model holds significant potential for drug development and infectious disease research.