A number of problems that affect cats are caused by tissue damage or loss from illness, injury, or aging. Adult stem cells maintain and heal tissues throughout an animal’s life and multi-potent stromal cells (ASCs) derived from fat have been found in fat tissue of many species. The cells can form different tissues like fat, bone, and cartilage, and cells from one animal can be used in another. Fat tissue is easily obtained during routine castration of male cats, so additional surgeries to get the tissue are not necessary. To date, cat ASCs are largely unexplored and an optimized and repeatable isolation technique is needed to provide cells for potential treatments. The best method to isolate feline ASCs and the potential for cell banking for treatment of ill and injured patients will be determined in this study. The results will significantly advance adult stromal cell applications to treat feline companions.