Biomimetic macroporous hydrogel scaffolds in a high-throughput screening format for cell-based assays

Area of Science:

• Biomaterials Science
• Tissue Engineering
• Cell Biology

Background:

• Macroporous hydrogels (MHs) are promising scaffolds for tissue engineering and cell-based assays.
• Integrating 3D cell culture with miniaturized screening offers enhanced experimental capabilities.

Purpose of the Study:

• To demonstrate the feasibility of combining 3D cell culture with a miniaturized screening format using functionalized MHs.
• To evaluate the impact of functionalized MHs on cancer cell aggregation and drug response.
• To assess the potential of this system for high-throughput toxicity testing.

Main Methods:

• MHs were fabricated using cryogelation and functionalized with type I collagen and an RGD peptide mimetic (abRGDm).
• Human colon cancer (HCT116), leukemia (KG-1), and fibroblast cells were cultured on MHs in a 96-minicolumn plate format.
• Scanning electron microscopy was used to visualize cell morphology and adhesion. Drug sensitivity assays were performed using cisplatin and Ara-C.

Main Results:

• Functionalized MHs promoted cancer cell aggregation and fibroblast adhesion, unlike plain MHs.
• Cells cultured as aggregates on functionalized MHs exhibited increased resistance to specific chemotherapeutic agents compared to single cells.
• HCT116 cells in 3D cultures on functionalized MHs were less sensitive to cisplatin than those in 2D cultures.

Conclusions:

• The developed system effectively combines 3D cell culture with a miniaturized format on functionalized MHs.
• Functionalized MHs influence cell behavior and drug response, creating a more physiologically relevant model.
• This platform holds potential for developing advanced in vitro models for high-throughput toxicity screening and drug development.