Porous chitosan microspheres as microcarriers for 3D cell culture

Highly porous chitosan microspheres (CSM) were prepared through emulsion-based thermally induced phase separation (TIPS) without using toxic crosslinkers and chemical porogenic agents other than ice. The CSM had an average diameter of ∼150 μm with interconnected pores varying from 20∼50 μm in size. Due to their excellent biocompatibility and unique porous structure, high-performance hepatocyte culture in three-dimensional (3D) space was achieved using the CSM as microcarriers, as cell growth also took place within the internal pores of the CSM, besides their external surface, and multidirectional cell–cell interactions were observed. Enhanced cellular activity and functions were obtained with the CSM microcarriers as compared with 2D cell culture. It is believed that these CSM microcarriers provide a promising platform for 3D cell culture in vitro.