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

Evidence from laboratory and animal studies provides groundwork for understanding mechanisms and potential before human studies continue.

• Level A · Stronger Clinical Evidence
• Level B · Emerging clinical evidence with positive signals
• Level C · Early human research exploring benefits
• Level D · Scientific groundwork from lab and animal studies
• Emerging · Emerging topic under active research

Abstract (original English)

Cell sheet engineering provides a scaffold-free strategy for fabricating cohesive tissue constructs, but challenges remain in maintaining structural integrity and mimicking complex tissue architectures. This study demonstrated perfluorodecalin-based liquid-liquid interfaces, known for their inertness and stability, as a simple, and efficient platform for fabricating cell sheets. Using single cells, spheroids, and their combination, we evaluated methods to enhance sheet formation. Single cells formed cohesive sheets at high densities (4 × 10 6 cells/well) but exhibited limited long-term stability due to nutrient constraints. Spheroids formed robust sheets at lower densities (2 × 10 6 cells/well), whereas higher densities impaired fusion. The mixed approach combined the advantages of both, improving uniformity, mechanical stability, and spheroid fusion, while mimicking muscle-like structures with vascular networks. Additionally, the cell sheets retained adipogenic and chondrogenic differentiation potential, highlighting their functional viability. These findings establish liquid interfaces as a practical and versatile platform for tissue engineering, regenerative medicine, and in vitro modeling.

Evidence level

Evidence from laboratory and animal studies provides groundwork for understanding mechanisms and potential before human studies continue.

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