On-Chip Fabrication of a Cell-Derived Extracellular Matrix Sheet

Area of Science:

• Biomaterials Engineering
• Cell Biology
• Microfluidics

Background:

• The extracellular matrix (ECM) is crucial for cell function, but integrating structural ECM into microfluidic devices remains challenging.
• Existing microfluidic systems often lack the complex biochemical and structural support provided by native ECM.

Purpose of the Study:

• To develop a microfluidic device incorporating a cell-derived ECM sheet for advanced cell culture applications.
• To evaluate the device's ability to mimic in vivo ECM microenvironments and influence cell behavior and mechanosensitivity.

Main Methods:

• A microfluidic device was fabricated with an ECM sheet derived from decellularized mouse embryonic fibroblasts (NIH/3T3).
• The ECM sheet's composition and architecture were characterized, confirming the presence of key ECM proteins and fibrous structures.
• Human umbilical vein endothelial cells (HUVECs) were cultured on the ECM sheet to assess cell morphology, adhesion, and response to fluid shear stress.

Main Results:

• The decellularization process preserved ECM components (collagens, fibronectin, laminin, elastin) and fibronectin architecture.
• HUVECs cultured on the ECM sheet displayed distinct morphologies and focal adhesion characteristics compared to controls.
• HUVECs on the ECM sheet showed enhanced mechanosensitivity, with focal adhesions and adherens junctions reorganizing in response to fluid shear stress, unlike control groups.

Conclusions:

• The developed microfluidic device effectively integrates a cell-derived ECM sheet, creating an in vivo-like microenvironment.
• This technology offers a valuable platform for studying cell-ECM interactions and mechanobiology in a controlled microfluidic setting.