The unique 1D structure of fibers offers intriguing attributes, including a high length‐to‐diameter ratio, miniatured size, light‐weight, and flexibility, making them suitable for various biomedical applications, such as health monitoring, disease treatment, and minimally invasive surgeries. However, traditional fiber devices, typically composed of rigid, dry, and non‐living materials, are intrinsically different from the soft, wet, and living essence of biological tissues, thereby posing grand challenges for long‐term, reliable, and seamless interfacing with biological systems. Hydrogel fibers have recently emerged as a promising candidate, in light of their similarity to biological tissues in mechanical, chemical and biological aspects, as well as distinct fiber geometry. In this review, a comprehensive overview of recent progress in hydrogel fibers‐based biointerfacing technology is provided. It thoroughly summarizes the manufacturing strategy and functional design, especially for hydrogel fibers with distinct optical and electron conductive performance, as well as responsiveness to triggers including thermal, magnetic field and ultrasonic wave, etc. Such unique attributes enable various biomedical applications, which are also examined in detail. Future challenges and potential directions, including biosafety, long‐term reliability, sterilization, multi‐modalities integration and intelligent therapeutic systems, are raised. This review will serve as a valuable resource for further advancement and implementation as next‐generation biointerfacing technology.