Historically important public inbred lines continue to play an important role in maize (Zea maysL.) improvement in many different breeding programs. Their continued use means they have undergone numerous seed increases in diverse programs since their original release. Our objective was to estimate the level of genetic diversity among and within inbred lines from different sources using SSR markers. We sampled six inbred lines (B73, CM105, Mo17, Oh43, W153R, and Wf9) obtained from 14 sources (breeding programs). The data were analyzed by analysis of molecular variance (AMOVA), genetic diversity statistics, and genetic distance (Dice's coefficient). Of the total variation observed in gene frequency, 87.8% was found among inbred lines, 7.6% among sources within inbred lines, and 4.6% within sources. Genotypes of identically named inbred lines from eight different sources differed slightly on the basis of 44 SSR loci. The mean genetic similarity between sources of the same inbred was greater than 85%. It can be concluded that although more diversity exists among these six inbred lines than within them, a small but significant amount of variation exists among seed sources within inbreds. This variation may have arisen through differences in seed maintenance, since we found no evidence to suggest high mutation rates or extensive outcrossing. The small but statistically significant level of variation raises concerns in germplasm conservation, mapping studies, marker development, and long-term recombinant inbred line development, especially when high resolution is desired.