Five members of Src family kinases (SFKs) namely Src, Fyn, Yes, Lyn and Lck are widely expressed in mammalian central nervous system. SFKs have been found to be involved in neuronal survival and neuronal cell death pathways; however the exact role of each of the members still remains unclear. SFK member Src kinase, is important for glial cell line-derived neurotrophic factor (GDNF) and thyroid hormone tri-iodothyronine (T3) mediated neurotrophic signalling pathways. Here we report the critical role of Src protein tyrosine kinase in supporting survival of cultured primary cortical neurons. Treatment with specific SFK inhibitor (SU6656) significantly reduced cell viability of cultured primary cortical neurons at day 7 (DIV7). Similar phenomenon was observed when neuronal Src was specifically knocked-down using specific short-hairpin RNA (shRNA), demonstrating its indispensable role in survival of primary cortical neurons. Both SFK inhibitor treatment and specific knock-down of Src expression was found to be associated with inhibition of Akt phosphorylation. Our data suggest that Src activity is crucial for neuronal cell survival and this activity is somewhat relate to neurotrophic Akt signalling pathway.