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

Submitting author affiliation:
University of Tokyo, Tokyo, Japan

Beilstein Arch. 2026, 202610. https://doi.org/10.3762/bxiv.2026.10.v1

Published 10 Mar 2026

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Abstract

Epithelial tissues form selective barriers essential for physiological homeostasis. Conventional in vitro models rely on solid substrate, which limit the physicochemical flexibility of the cellular microenvironment. Here, we introduce a microfluidic platform in which a collagen-coated liquid-liquid interface formed between perfluorocarbon (FC-43) and culture medium serves as a substrate for epithelial cell adhesion. By culturing Caco-2 cells in the device, we show that the liquid interface supports cell attachment and the formation of near-confluent monolayers. Immunofluorescence observation reveals the development of tight junctions and organized actin cytoskeletons, indicating early-stage epithelial maturation. Our new microfluidic system enables the formation of stable liquid-liquid interfaces that serve as viable and flexible substrates for epithelial cell culture, offering new opportunities for multiphase microfluidic models of epithelial barriers.

Keywords: gut-on-a-chip; liquid-liquid interface; collagen; Caco-2; FC-43

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Kuriu, S.; Kim, S. H. Beilstein Arch. 2026, 202610. doi:10.3762/bxiv.2026.10.v1

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