In vitro biomimetic platforms featuring a perfusion system and 3D spheroid culture promote the construction of tissue-engineered corneal endothelial layers

This study demonstrated that using a perfusion culture bioreactor and 3D spheroid culture improved the expansion and construction of tissue-engineered corneal endothelial cell (CEC) layers in vitro. Perfusion-derived CECs showed increased proliferation, enhanced cell-cell contact, and numerous surface microvilli compared to static culture. The spheroid culture down-regulated proliferation and EMT-related markers while up-regulating the CEC marker ATP1A1. Additionally, combining the perfusion system with spheroid culture on decellularized corneal scaffolds and collagen sheets facilitated the formation of CEC monolayers and extracellular matrix. The study confirmed that a CEC spheroid culture on a curved collagen sheet under controlled intraocular pressure could generate a CEC monolayer, highlighting the potential of these methods for tissue engineering applications.