Neural stem and progenitor cells (NSC/NPC) from the subventricular zone (SVZ) are able to generate all neural cell types and can be cultured as free-floating aggregates called neurospheres (NS). NS cultures are useful tools to model the cellular processes that take place during endogenous repairment that occur in the central nervous system (CNS) in a sequential and orderly manner.
In the present work, we first developed NS cultures to evaluate the effect of the demyelinating agent Cuprizone (CPZ) on NSC/NPC behavior. CPZ treatment of NS cultures revealed its direct effect on NSC/NPCs proliferation and differentiation mechanisms. The finding that cell migration was also affected by CPZ, led us to investigate extracellular matrix (ECM) involvement in the control of cell migration from NS. Hyaluronic acid (HA) is a linear non-sulfated glycosaminoglycan found in the ECM that acts as structural support and exerts regulatory functions in the SVZ over development and regeneration processes. We used the inhibitor of HA synthesis 4-methylumbelliferone (4-MU) to evaluate HA requirement for cell migration in NS cultures. HA depletion affected NSC/NPC proliferation, as it diminished NS size at doses higher than 125µM 4-MU, and also reduced the number of migratory cells detached from adherent NS. These findings reinforce HA requirements for the control of NSC/NPC proliferation and migration and could help to develop new strategies to improve CNS regeneration.