Gang Zhao, Yue Cheng, Conghui Tian, Guangzhu Shang
Recent advances in microencapsulation for cryopreservation have transformed cell preservation through innovative hydrogel-based strategies. By leveraging the ice-inhibiting properties of three-dimensional hydrogel networks, this approach significantly reduces reliance on toxic cryoprotectants while maintaining high cell viability. The technology has demonstrated remarkable success across diverse cell types, from human mesenchymal stem cells and erythrocytes to murine reproductive tissues. This chapter presents a comprehensive protocol using mouse follicles as a model system, detailing: (1) mechanical follicle isolation, (2) centrifugal microfluidic encapsulation in alginate hydrogels, (3) synthesis of magnetothermal nanoparticles, and (4) optimized vitrification/rewarming processes. The integrated methodology combines the protective benefits of physical encapsulation with advanced nanowarming techniques, establishing a robust platform for low-toxicity, high-efficiency cryopreservation.