This study investigates the use of bioprinted skeletal muscle constructs to facilitate neurite growth from isolated dorsal root ganglia. By embedding neurotrophic factors in microspheres within the constructs, the research aims to create a controlled environment for neuronal development.
Take bioprinted skeletal muscle constructs.
These constructs contain fibrin hydrogel pillars with embedded muscle fibers and microspheres.
The microsphere is a porous polymer structure enclosing neurotrophic factors.
Take dorsal root ganglia isolated from chick embryos.
Place one ganglion on each construct and incubate.
The neurotrophic factors are released from the microspheres at a controlled physiological concentration, facilitating the sprouting of projections from the neurons called neurites.
Add a fixative to fix the constructs.
Wash with a buffer and add a blocking solution to prevent nonspecific antibody binding.
Incubate with the primary antibodies, which bind specifically to the neurite.
Rinse with buffer to remove unbound antibodies.
Add fluorescently tagged secondary antibodies that bind to the primary antibodies.
Wash with buffer to remove excess antibodies.
Mount the construct using mounting media containing a nuclear dye and a coverslip.
Under a confocal microscope, visualize the neurite growth.
繁體中文
