Anchored Cell Sheet Engineering: A Novel Scaffold-Free Platform for in vitro Modeling

This work introduces a next-generation scaffold-free tissue engineering platform called Anchored Cell Sheet Engineering (ACSE), enabling fabrication of self-assembled 3D tissue constructs from enzyme-free cell sheets. The system integrates patterned PDMS membranes and flexible Ecoflex-based pillar anchors to control tissue form, ECM deposition, and remodeling. The platform demonstrated the creation of muscle-fiber-like and lung-epithelial constructs without exogenous scaffolds. Decellularized cell-sheet constructs (CSdECM) were shown to act as tunable ECM-rich scaffolds and coatings for improving 2D and 3D culture fidelity. This versatile, low-cost system allows modeling of complex assembloids and interlinked tissues, advancing in vitro mechanobiology and regenerative research.