A Biomimetic Buffering Hydrogel Scaffold for Long‐Term Culture of Patient‐Derived Tumor Organoids

Patient-derived cancer organoids have emerged as a promising in vitro model for fundamental cancer research and drug screening for therapeutic cancer treatment. Yet, while the inherent acidification of the tumor environment in vivo is controlled at a particular level, hydrogel scaffolds used for organoid culture lack this ability and their pH falls outside the physiologically relevant range. The excessive acidification can also lead to the degradation of pH-sensitive hydrogel scaffolds during long-term organoid culture, thus changing the mechanical properties of the organoid microenvironment. Here, we report a biomimetic fibrous hydrogel with built-in buffering capacity, which enables control of the local acidification of the organoid environment to maintain its mechanical and structural stability. The hydrogel is formed from aldehyde-functionalized cellulose nanocrystals carrying histidine buffering molecules, and gelatin. During long-term organoid culture, the hydrogel maintained the pH in the physiologically relevant range, while maintaining network integrity and mechanical properties. The organoids grown in this hydrogel exhibited enhanced proliferative activity of cancer cells, thus reflecting a more homeostatic tumor-like niche. This work shows that introducing a buffering functionality into the hydrogel scaffold enables significantly improved support for long-term culture of patient-derived breast cancer organoids under physiologically relevant conditions.