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Interface-engineered Caco-2 cell culture on a collagen-coated liquid-liquid interface in a microfluidic device.

Satoru Kuriu¹, Soo Hyeon Kim¹

¹Institute of Industrial Science, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8505 JAPAN.

Beilstein Journal of Nanotechnology

|June 17, 2026

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Researchers developed a novel microfluidic platform using a liquid-liquid interface for culturing epithelial cells. This flexible substrate supports cell growth and maturation, advancing in vitro barrier models.

Area of Science:

Biomedical Engineering
Cell Biology
Microfluidics

Background:

Epithelial tissues are crucial selective barriers for physiological homeostasis.
Conventional in vitro models using solid substrates have limitations in physicochemical flexibility.
Developing advanced cell culture models is essential for studying epithelial barrier function.

Purpose of the Study:

To introduce a novel microfluidic platform utilizing a liquid-liquid interface as a substrate for epithelial cell culture.
To assess the viability and suitability of this liquid interface for supporting epithelial cell adhesion, monolayer formation, and maturation.
To explore the potential of this system for creating advanced multiphase microfluidic models of epithelial barriers.

Main Methods:

A microfluidic platform was designed with a collagen-coated liquid-liquid interface between perfluorocarbon (FC-43) and culture medium.

Keywords:

Caco-2 FC-43 collagen gut-on-a-chip liquid–liquid interface

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Caco-2 cells were cultured on this liquid interface within the microfluidic device.

Immunofluorescence microscopy was employed to observe cell attachment, monolayer formation, tight junction development, and actin cytoskeleton organization.

Main Results:

The liquid-liquid interface successfully supported Caco-2 cell attachment and the formation of stable epithelial monolayers.
Immunofluorescence analysis confirmed the development of tight junctions and organized actin cytoskeletons, indicative of early epithelial maturation.
The microfluidic system demonstrated the ability to form stable liquid interfaces suitable for cell culture.

Conclusions:

A novel microfluidic system with a stable liquid-liquid interface provides a viable and flexible substrate for epithelial cell culture.
This platform facilitates the study of epithelial barrier formation and maturation in vitro.
The system offers new possibilities for developing advanced multiphase microfluidic models relevant to physiological research.