Enhanced cell sheet engineering through combination of single cells and spheroids on liquid interface using...

Area of Science:

• Biomaterials Science
• Tissue Engineering
• Regenerative Medicine

Background:

• Cell sheet engineering offers scaffold-free tissue fabrication but faces challenges in structural integrity and mimicking complex architectures.
• Perfluorodecalin-based liquid-liquid interfaces provide an inert and stable platform for cell culture.

Purpose of the Study:

• To evaluate perfluorodecalin liquid interfaces as a platform for fabricating robust and functional cell sheets.
• To compare cell sheet formation using single cells, spheroids, and a combination of both.

Main Methods:

• Fabrication of cell sheets on perfluorodecalin liquid-liquid interfaces using single cells and/or spheroids.
• Assessment of cell sheet formation, uniformity, mechanical stability, and fusion at varying cell densities.
• Evaluation of the differentiation potential (adipogenic and chondrogenic) of the fabricated cell sheets.

Main Results:

• Single cells formed cohesive sheets at high densities but showed limited stability.
• Spheroids formed robust sheets at lower densities, with higher densities hindering fusion.
• A mixed approach (single cells and spheroids) improved uniformity, mechanical stability, and fusion, creating muscle-like structures with vascular networks.
• Cell sheets retained adipogenic and chondrogenic differentiation potential.

Conclusions:

• Perfluorodecalin liquid interfaces offer a simple, efficient, and versatile platform for scaffold-free cell sheet fabrication.
• This approach advances tissue engineering, regenerative medicine, and the development of in vitro models.
• The method successfully mimics complex tissue architectures and preserves cell functionality.