Seasonal, pandemic, and avian influenza virus infections may be associated with central nervous system pathology, albeit with varying frequency and different mechanisms. Here, we demonstrate that differentiated human astrocytic (T98G) and neuronal (SH-SY5Y) cells can be infected by the recently emerging avian influenza A H7N9 viruses. Much like the low pathogenic pandemic H1N1 virus (pdmH1N1), H7N9 can infect and cause cytopathic effects in human astrocytes and neuronal cells, however only H7N9 produces infectious progeny viruses in human neuronal cells. Neither of these viral strains can generate infectious progeny virus in human astrocytes despite replication of viral genome was detected. Furthermore, H7N9 virus triggered high pro-inflammatory cytokine expression, whilst pdmH1N1 virus induced only low cytokine expression in either brain cell type. The experimental finding here is the first data to demonstrate that the highly pathogenic avian H7N9 virus can infect, replicate, induce cytokine upregulation and cause cytopathic effects in human astrocytes and neuronal cells, and thus may potentially lead to profound central nervous system injury. Observation for neurological problems due to H7N9 virus infection deserves further attention when managing these patients.