To detect cell cross-contamination and verify the origin of the cells of an artificial organ, the sensitive isoenzyme assay was chosen to monitor the quality test of cell-based devices. Authoritative cell evaluation of artificial skin products has been established in this study. Human and porcine cell suspensions with total cell counts of between 1 × 10 5 and 4 × 10 6 were individually tested to determine the activity of isoenzymes. Human fibroblast, mixed with 1% to 100% of porcine fibroblast, could be significantly distinguished in the isoenzyme assay. Based on the glucose-6-phosphophatedehydrogenase analysis, the human fibroblast tested in this study belonged to the B type human cells. Lactate dehydrogenase (LD), malate dehydrogenase (MD) and mannose phosphate isomerase isoenzyme (MPI) activities obviously revealed that a different pattern corresponds to the percentage of human and porcine cell mixtures. The discriminatory limit of MPI, LD and MD activity can reach up to 1% of sensitivity of the isoenzyme analysis. This sensitive isoenzyme analysis method allows us to routinely test cellular biomaterials whether interspecies cell line cross-contamination has occurred in the development of artificial organs.