Multiplex recreation of human intestinal morphogenesis on a multi-well insert platform by basolateral convective flow

Area of Science:

• Biotechnology
• Tissue Engineering
• Gastroenterology

Background:

• Current in vitro models often fail to replicate the complex 3D microarchitecture of the human intestinal epithelium.
• Developing physiologically relevant models is crucial for accurate drug screening and disease modeling.

Purpose of the Study:

• To develop a multiplex culture system for simultaneous, in vitro recreation of multiple 3D human intestinal epithelial microarchitectures.
• To establish a scalable and reproducible platform for studying intestinal barrier function and drug interactions.

Main Methods:

• Utilized the basolateral convective flow-generating multi-well insert platform (BASIN) with nano-porous inserts.
• Applied controllable convective flow to recreate biomimetic morphogen gradients, inducing intestinal morphogenesis.
• Employed Caco-2 human intestinal epithelium and characterized cytodifferentiation, proliferation, and barrier function.

Main Results:

• Successfully regenerated a 3D villi-like intestinal microstructure with high barrier function and minimal variation.
• Demonstrated enhanced cytodifferentiation and proliferation using specific cell markers (villin, Ki67).
• Confirmed reproducibility, robustness, and scalability for transport and efflux studies, and validated with dextran sodium sulfate.

Conclusions:

• The BASIN platform provides a novel, multiplex, and scalable in vitro model of the human intestinal epithelium.
• This system accurately mimics intestinal morphology and barrier function, suitable for biochemical, pharmaceutical, and toxicological studies.
• The BASIN platform facilitates the construction of leaky-gut models and offers a valuable tool for basic and applied research.