Ames was established in 1864 to serve the Cedar Rapids and Missouri River Railroad. Cynthia Duff, an early Ames settler, was instrumental in helping to secure land for the railroad.

The railroad president, John Blair, named the town after his friend Oakes Ames, a railroad promoter and congressman from Massachusetts, who never lived in Ames.

In 1870, the town was incorporated with a population of 844. It was platted by Charles Irish, a railroad engineer. Ames consisted of 12 blocks and extended from the railroad right-of-way north to Ninth Street and was bounded on the east by Duff Avenue and on the west by Burnett Avenue.

That year also began the establishment of the first of the state and federal institutions which have influenced Ames' growth. Iowa Agricultural College and Model Farm was established as a Land-Grant College. The first term enrollment included 77 men and 16 women. Today, as Iowa State University of Science and Technology (ISU), it serves over 27,000 students from all over the world.

The Iowa Department of Transportation, formerly known as the Iowa Highway Commission, was created in 1913.

Ames also has two Partner City agreements.
- Enzan City, Japan, was established by resolution number 93-353 as a partner city on September 14, 1993.
- Frydek-Mistek, Czech Republic, was established as a partner city on June 11, 1996, by resolution number 96-254.

The current Ames population is estimated at 52,319 (2004).