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Intestinal Epithelium Tubules on a Chip
Kinga Kosim1, Iris Schilt1, Henriëtte L Lanz1
1Mimetas BV, Leiden, The Netherlands.
Insights
This study introduces a high-throughput microfluidic platform for culturing 3D intestinal tubules. This method enhances in vitro studies of epithelial barrier properties, offering a membrane-free approach for disease modeling and drug transport research.
Area of Science:
- Biomedical Engineering
- Cell Biology
- Physiology
Background:
- Epithelial barrier properties are crucial for disease modeling, drug transport, toxicology, and developmental/regenerative biology.
- Current in vitro methods use static cell cultures on artificial membranes, limiting physiological relevance.
- Advancements in microfluidics and organ-on-a-chip technologies enable more complex in vitro models.
Purpose of the Study:
- To describe protocols for culturing 3D intestinal tubules in a microfluidic, high-throughput format.
- To enable assessment of epithelial tubule permeability and marker expression.
- To facilitate the adoption of microfluidic techniques by non-specialized users.
Main Methods:
- Culture of 3D intestinal tubules directly against extracellular matrix (ECM) under flow using microfluidic chips.
- High-throughput format with 40 independent microfluidic chips in a microtiter plate.
- Parallel analysis of epithelial tubules using high-content microscopy.
Main Results:
- Successful establishment of 3D intestinal tubule cultures in a microfluidic platform.
- Demonstrated capability to assess permeability and marker expression in parallel.
- Developed protocols suitable for routine application by non-specialized researchers.
Conclusions:
- Microfluidic organ-on-a-chip technology provides a membrane-free, physiologically relevant in vitro model for studying epithelial barriers.
- The described high-throughput platform simplifies the application of advanced microfluidic techniques.
- This approach has broad implications for disease modeling, drug development, and regenerative medicine.
Abstract:
The study of epithelial barrier properties in the human body is of paramount interest to a range of disciplines, including disease modeling, drug transport studies, toxicology, developmental biology, and regenerative biology. Current day in vitro studies largely rely on growing epithelial cells in a static environment on membrane cell culture inserts. With the advancement of microfluidic and organ-on-a-chip techniques it became possible to culture 3D intestinal tubules directly against an extracellular matrix (ECM) under flow and without the need for artificial membranes. Here we describe detailed protocols for culturing epithelial tubules in a high-throughput format, assessing their permeability and marker expression. The platform harbors 40 independent microfluidic chips in a microtiter plate format. The resulting 40 epithelial tubules are analyzed in parallel using a high-content microscopy. Protocols described here allow for adoption and routine application of microfluidic techniques by nonspecialized end-users.
