The invention belongs to the cross technical field of
cell engineering, nano-drugs and biological manufacturing, and particularly discloses a
drug-loaded
vesicle based on denucleated cells as well as a preparation method and application of the
drug-loaded
vesicle. According to the invention, a
cell suspension and a
cytoskeleton relaxant are co-incubated to relax an
actin skeleton and weaken
nucleoplasm connection; then loading the
treated cell suspension on a multi-layer discontinuous
density gradient centrifugal medium, realizing
physical separation of
cell nucleuses and
cytoplasm through high-speed
centrifugation according to
buoyancy density difference, and collecting components of a specific interface to obtain high-purity denucleated cells with a complete
membrane structure; then co-incubating the denucleated cells and ROS
response type lipidosome (co-carrying therapeutic siRNA and a sound-sensitive agent Ce6) prepared in advance, so that the lipidosome is wrapped or anchored by a denucleated
cell membrane; finally, the composite
system is subjected to
extrusion treatment through a microporous membrane, the
drug-loaded vesicles uniform in particle size and stable in structure are obtained, and the drug-loaded vesicles are suitable for various application scenes such as anti-tumor treatment and
RNA vaccine delivery.