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In 1969, Nobel Prize-winning scientist Peter Medawar predicted that scientific innovation would someday fully replace the use of animals in biomedical research. Medawar correctly forecast the leveling off and subsequent decline in animal use in the last quarter of the 20th century – a period of remarkable innovation in the life sciences. A 2007 report by the U.S. National Academy of Sciences, Toxicity Testing in the 21st Century, proposed a strategy that is likely to replace all routine animal use in toxicology with innovative methods within one to two decades. Replacing animal use throughout biomedical research is more challenging given its diverse nature and larger scale. Nonetheless, full replacement is a goal worth pursuing for a host of reasons. This presentation will outline these reasons, discuss animal use trends, review current replacement initiatives and challenges, and call for coordinated, targeted, and sustained efforts to fully replace animals in research and testing.