We studied the inhibitory effect of urinary macromolecular substances (UMM), more than 10,000 mol. wt., on aggregation of calcium oxalate crystals in a crystal-seed system and a non-crystal-seed system. In both assay systems, UMM from non-stone-farmers showed a marked inhibitory activity on crystal aggregation, whereas UMM from recurrent stone-formers showed a weak inhibitory activity. The distinction between the two types of UMM was greater in the non-crystal-seed system. The macromolecular inhibitors seemed to be composed of proteins or protein-complexes inasmuch as approximately 70 to 90 per cent of the macromolecular inhibitory activity was destroyed by protein digestion with pronase. Urinary acid glycosaminoglycans, which have been considered possible inhibitors, played a small role in the inhibition of crystal aggregation.