Reactive astrocytes play important pathophysiological roles in various brain diseases in a context-dependent manner, and they have thus gained significant attention as a target for development of new drugs. Therefore, we aimed to characterize human astrocytes/conditionally immortalized, clone 35 (HASTR/ci35) as an in vitro reactive human astrocyte model. The results of quantitative PCR and RNA-sequencing showed that, upon exposure to pro-inflammatory cytokines, HASTR/ci35 cells exhibited significant up-regulation of mRNA levels of various inflammation associated genes, such as interleukin-6, intercellular adhesion molecule 1, and pentraxin 3. The results of the protein array confirmed their induction at protein levels. Qualitatively, the response profile of HASTR/ci35 cells appeared to be similar to that observed in human primary astrocytes. To summarize, HASTR/ci35 cells show an unequivocal reactive response to inflammatory stimuli, indicating that they can serve as a human reactive astrocyte model. While further characterization is currently underway, it can be expected that they have a considerable potential to be used in reactive astrocyte-targeted drug development studies.