Figure 3.

Overview of BM nerve fiber isolation, culturing, and neurosphere transplantation. Adult mice were sacrificed, and the femur, tibia, and pelvis dissected. The BM plug was liberated by flushing the medullary cavity with PBS, then gently triturated to free the nerve fibers from the stroma. Nerve fibers were identified by fluorescent microscopy and transferred to neuroproliferation media for culture by pipette aspiration. For in vivo transplantation experiments, neurospheres were surgically implanted into the colon or colorectum via injection with a microliter syringe.