We describe an innovative method of culturing a three-dimensional cell mass (3DCM) of human adipose-derived stem cells (hASCs) in the xeno-free (XF) condition for the treatment of severe ischemic diseases. The majority of the mice injected with XF-3DCMs exhibited limb salvaging and displayed similar blood perfusion compared to normal limbs. In vivo tumorigenicity and toxicity analysis showed that XF-3DCMs did not transform into tumor cells and induce toxicity, respectively. Our results strongly suggest that XF-3DCMs can be effectively used for therapeutic applications and eliminate immunological reaction of animal-derived material contamination.
Keywords: cell therapy; human adipose-derived stem cells; three-dimensional stem cell mass; toxicity test; tumorigenicity test; xeno-free media.