Large-scale three-dimensional (3D) cell culture plays a critical role in vaccine production, regenerative medicine, and drug development. However, the 3D carriers currently used for large-scale cell culture are limited by high production costs, poor reusability, and a lack of multifunctionality. In this study, we develop a magnetic gelatin microcarrier (GM) platform by combining amino-modified gelatin (AmGel) and vinyl-modified Fe3O4 nanoparticles (MAFe3O4) using microfluidic technology. This system not only supports efficient cell adhesion and proliferation but also enables magnetic-assisted recovery for repeated use, significantly outperforming commercial microcarriers in cell culture efficiency. Furthermore, the magnetothermal properties of GMs facilitate the in situ induction of immunogenic cell death (ICD) under an alternating magnetic field (AMF), generating a whole tumor cell vaccine with the GM and the ICD fragments. Upon injection into living mice with or without tumors, the GM-based vaccine combined with localized AMF-induced hyperthermia effectively promotes dendritic cell recruitment, enhances immune responses, and effectively inhibits tumor growth. Therefore, this study provides proof-of-concept for a multifunctional platform using GMs for large-scale cell culture, immunotherapy, and cancer vaccine development.