Effect of culture conditions on the phenotype of adult neural stem cells from the SVZ

PIANTANIDA A. P.1, MATHIEU P. A.1 AND PITOSSI F.1

Huerta Grande - Córdoba

Congreso; II Reunión Conjunta de Neurociencias; 2010

              Adult neural stem cells (aNSC) from the subventricular zone (SVZ) generate new neurons throughout life. They can be cultured as floating neurospheres, and can generate secondary spheres upon mechanical dissociation. Culture conditions affect aNSC differentiation rate. For example, TGF-β1 is pro-neurogenic on hippocampal aNSCs. In order to consider these cells for potential regenerative therapies,is essential to develop efficient and reproducible culture protocols.             We aimed to characterize the effect of different culture conditions on cell phenotypes of aNSC cultures from SVZ and to study whether TGF-β1 has a pro-neurogenic role. We generated neurospheres from adult rat SVZ, and performed immunocytochemistry with markers for aNSCs (Nestin), astrocytes (GFAP), and neurons (Tuj). TGF-β1 was added to these cultures for 5 days.             Cultures maintained without mechanical dissociation showed a higher Nestin (92%)- and a lower Tuj (0,4%)-positive fraction when grown with EGF, while the GFAP-positive fraction was not altered. When dissociated, cultures showed an initial higher fraction of Tuj-positive cells. Upon EGF treatment, the Nestin-positive cell fraction was also increased, while Tuj- and GFAP-fractions did not vary. TGF-β1 treatment of dissociated cultures increased the Tuj-positive cell fraction.             Thus, EGF increases aNSCs fraction in these cultures, while mechanical dissociation and TGF-β1 increase neuronal phenotypes. These factors may be powerful tools to increase a desired cell phenotype within aNSC populations.