Transparent 3-Layered Bacterial Nanocellulose as a Multicompartment and Biomimetic Scaffold for Co-Culturing Cells

Area of Science:

• Biomaterials Science
• Cell Biology
• Cancer Research

Background:

• Three-dimensional (3D) cell culture models offer a more physiologically relevant microenvironment than traditional 2D cultures.
• Existing 3D models vary significantly in complexity and cost.
• Simulating the tumor microenvironment (TME) is crucial for understanding cancer progression and developing therapies.

Purpose of the Study:

• To introduce and characterize a novel translucent, multi-compartmentalized, stacked, multilayered nanocellulose scaffold.
• To evaluate the scaffold's potential for co-culturing multiple cell types, specifically simulating the TME.
• To assess the biological relevance of the scaffold by analyzing gene expression in co-cultured cells.

Main Methods:

• Fabrication and characterization of a bacterial nanocellulose (BNC) scaffold with distinct layers.
• Co-culturing MDA-MB-231 breast cancer cells with tumor-associated fibroblasts (BC-CAFs) and M2 macrophages within the scaffold.
• Assessing cell viability, metabolic activity, cell invasion using confocal microscopy, and gene expression analysis (E-cadherin).

Main Results:

• Cells remained viable and metabolically active for up to 15 days in the scaffold.
• No signs of cell invasion were observed, but BC-CAFs and cancer cells co-localized within layers.
• Cancer cells in the scaffold showed a 20% decrease in E-cadherin expression compared to 2D cultures.

Conclusions:

• The novel nanocellulose scaffold provides a viable and functional platform for simulating the TME.
• The observed downregulation of E-cadherin suggests the scaffold better recapitulates in vivo TME complexity than 2D cultures.
• This scaffold holds promise for advancing cancer research and drug development by offering a more accurate model of the tumor microenvironment.