There are compelling reasons to develop a tissue-engineered mitral valve, but this endeavor has not received the same attention as tissue engineering strategies for the semilunar valves. Challenges in regenerating a mitral valve include recapitulating the complex heterogeneity in terms of anatomy (differently sized leaflets, numerous chordae), extracellular matrix composition, biomechanical behavior, valvular interstitial cell and endothelial cell phenotypes, and interior vasculature and innervation. It will also be essential to restore the functional relationships between the native mitral valve and left ventricle. A growing amount of information relevant to tissue engineering a mitral valve has been recently collected through investigations of cell mechanobiology and collagen organization. It is hoped that the development of tissue-engineered mitral valves can build on knowledge derived from engineering semilunar valves, but the mitral valve will present its own unique challenges as investigators move toward a first-generation prototype.