Fibronectin in Layer-by-Layer Assembled Films Switches Tumor Cells between 2D and 3D Morphology

Area of Science:

• Biomedical Engineering
• Cell Biology
• Materials Science

Background:

• Three-dimensional (3D) tumor cell morphology and coculture with endothelial cells offer valuable in vitro models for pharmaceutical development and studying cell-cell interactions.
• Understanding differential cell adhesion is crucial for engineering complex cellular microenvironments.

Purpose of the Study:

• To engineer micropatterned heterotypic cocultures of HepG2 tumor cells and endothelial cells.
• To investigate the role of fibronectin and poly(allylamine hydrochloride) in modulating cell adhesion and 3D morphology.
• To explore the impact of transglutaminase treatment on tumor cell morphogenesis.

Main Methods:

• Utilized microfluidics to create micropatterned cocultures based on differential cell adhesion.
• Investigated HepG2 cell and endothelial cell adhesion to fibronectin-coated surfaces, alone and with poly(allylamine hydrochloride).
• Assessed the effects of a hybrid extracellular matrix and transglutaminase treatment on cell behavior and 3D morphogenesis.

Main Results:

• HepG2 cells exhibited distinct adhesion patterns compared to endothelial cells on fibronectin-based substrates.
• Engineered cocultures demonstrated spatially encoded and physiologically relevant cell functions.
• The hybrid extracellular matrix synergistically modulated HepG2 cell protrusion, migration, and 3D morphology.
• Transglutaminase treatment inhibited tumor cell 3D morphogenesis.

Conclusions:

• Differential cell adhesion to fibronectin can be exploited to engineer specific tumor cell coculture architectures.
• The developed hybrid materials and microfluidic techniques provide a platform for advanced in vitro modeling of tumor microenvironments.
• Findings contribute to the development of novel pharmaceutical strategies targeting tumor cell behavior and interactions.