Transient receptor potential A1 (TRPA1) is a cold sensitive cation channel， which could also be activated by various pungent compounds. As a transduction channel in a number of sensory modalities, TRPA1 expressing in heterogonous systems serves to provide great convenience in pharmacological analysis and functional investigation. Due to cellular toxicity, establishment of stable TRPA1 cell line has always been challenging. Nevertheless, the first stable human embryonic kidney (HEK-293) cell line with un-controlled expression of TRPA1 was successfully established. It was also confirmed that this stable cell line retained TRPA1 expression for more than 25 passages in culture. The functional analysis of the cell response verified the stability and specificity of this novel recombinant TRPA1 cell line. Altogether, the data indicated this TRPA1-HEK cell line would be a useful tool for functional analysis of TRPA1 and for the development of high throughput screening (HTS) compatible assay in the effort to identify TRPA1 modulators.