Hyaluronic acid depletion effects on neurosphere cultures

MOLINARI Y.; PIBUEL M.; POODTS D; DÍAZ M.; LOMPARDÍA S.; SILVESTROFF L; FRANCO P

CÓRDOBA

Congreso; XXXIV Reunión Anual de la Sociedad Argentina de Investigación en Neurociencias (SAN); 2019

Sociedad Argentina de Investigación en Neurociencias (SAN)

Hyaluronic acid (HA) is a linear non-sulfated glycosaminoglycan found in the extracellular matrix that acts as structural support and exerts regulatory functions in several tissues. In the brain, HA is critical for the maintenance of subventicular zone (SVZ) architecture and acts as a neural stem cell and progenitor cell (NSC/NPC) niche during CNS development and regeneration. HA dysregulation has been previously associated to remyelination failure in animal models of demyelination.In the present work we isolated NSC/NPC from periventricular mouse brain tissue and cultured as neurospheres (NS) in order to study in vitro HA requirements for NSC/NPC biological functions.NS cultures were first characterized to confirm cell multipotency. We evaluated the effects of 4-Methylumbelliferone (4-MU), a coumarin derivative which inhibits HA synthesis, throughout NS culture progression. Floating NS were generated at all doses of 4-MU used. NS size significantly decreased at doses higher than 125µM 4-MU, suggesting that HA synthesis inhibition affects NSC/NPC proliferation and correlating with a significant decrease in 4-MU-treated culture metabolic activity. Adhered NS treated with 4-MU showed a reduced number of migratory cells and inhibition in the elongation of GFAP+ processes from the NS surface.These findings reinforce HA requirements for the control of NSC/NPC proliferation and migration and could help develop new strategies to improve CNS regeneration.