The Museum was founded in 1944 by a group of historically minded citizens who had acquired the mule-drawn streetcar "Bellefontaine."
In 1948, the Transport Museum Association (TMA) incorporated as a non-profit educational organization to better serve the financial and volunteer needs of the Museum. Land was acquired on Barrett Station in St. Louis County, along the right-of-way of the historic Missouri Pacific Railroad, and over the years, the mule-drawn streetcar was joined by hundreds of other significant exhibits.
On September 1, 1979, the St. Louis County Department of Parks and Recreation formally assumed the operation and development of the Museum, accepting it as a gift from the original founders in February 1984. Today, its mission is 'to be the leader in protecting and interpreting North America's transportation heritage.'