Dual-enzymatically crosslinked hyaluronic acid hydrogel as a long-time 3D stem cell culture system.

Stem cell based tissue engineering shows enormous potentials in regenerative medicine. Three-dimensional (3D) stem cell culture is the most basic part for tissue engineering. However, a perfect scaffold for high efficient 3D cell culture is still limited currently. Herein, a new hyaluronic acid hydrogel dual-enzymatically crosslinked by horseradish peroxidase (HRP) and choline oxidase (ChOx) is developed as a 3D stem cell culture system. This hydrogel possesses a superior stability over 2 months, controllable biodegradability with hyaluronidases, high swelling ratio exceeding 6000%, and excellent cytocompatibility in vitro and biocompatibility in vivo. More importantly, a long-time and highly cellular activity 3D culture of Bone marrow-derived MSC (BMSC) over 20 days was achieved in vitro. All these encouraging results highlight the great prospect of this new hydrogel for 3D culture and tissue engineering.