Light‐Activated Isolation of High‐Quality Mitochondria for Therapeutic Transplantation

ABSTRACT Artificial mitochondrial transplantation (AMT) holds great promise for reprogramming cellular metabolism and restoring cell function. Its clinical translation, however, relies on access to mitochondria that are both of high purity and metabolically active, requirements that current isolation techniques struggle to meet. Conventional differential centrifugation (DC) method yields heterogeneous and low‐activity mitochondria, whereas magnetic bead (MB)‐based immuno‐isolation leaves non‐biodegradable beads permanently attached. Herein, we present a Light‐Activated Mitochondrial Isolation ( LAMI ) platform comprising programmable mitochondria‐targeting MBs and a photo‐responsive release mechanism for the selective, efficient, and non‐destructive extraction of high‐quality mitochondria. LAMI employs magnetic nanoparticles decorated with a branched, modular probe architecture that supports systematic variation in mitochondria‐targeting ligand type, ligand density, and optical tracking elements. Incorporation of a photo‐cleavable linker allows on‐demand, mild, and reagent‐free release of captured mitochondria. Compared with DC method, LAMI produces mitochondria with markedly improved purity, structural integrity, and functionality. In an ischemia‐reperfusion injury (IRI) model, LAMI ‐isolated mitochondria‐based AMT exhibits superior therapeutic performance. Together, LAMI provides a non‐destructive, efficient, and versatile mitochondrial isolation strategy that overcomes long‐standing limitations of current methods, offering a robust platform to advance AMT and its future biomedical applications.