Fuxian Liu, Zheng Cai, Yuanhuan Dai, Xuan Ma, Rui Jiang, Jiamei He, Ancheng Lu, Renjie Chang, Junzi Wu
Organoids are high-fidelity in vitro models with substantial potential in biomedicine. However, traditional animal-derived matrix gels contain undefined components and have unpredictable mechanical properties, which greatly hinder the standardization and functional maturation of organoids. In recent years, nanofibers have emerged as an ideal substrate for organoid cultivation because of their structural similarity to natural extracellular matrices and their easily tunable properties. Accordingly, this article systematically reviews the synergistic effects of nanofibers across chemical, physical, and biological dimensions, along with research progress in transitioning organoids from passive culture to active engineering. Furthermore, the review summarizes current challenges and identifies future directions for development. The aim is to provide a systematic engineering design framework for next-generation biomimetic organoid culture matrices and promote their advancement toward standardization and clinical translation.