Under the program Ford self-driving cars, with safety drivers, will be deployed on the Lyft network in Miami later this year and in Austin starting in 2022.
The companies are planning to deploy at least 1,000 autonomous vehicles on the Lyft network in various locations over the next five years.
Lyft will take a stake in the autonomous platform Argo AI, which will supply the technology for the venture.
"This collaboration marks the first time all the pieces of the autonomous vehicle puzzle have come together this way," Lyft co-founder and chief executive Logan Green said.
"Each company brings the scale, knowledge and capability in their area of expertise that is necessary to make autonomous ride-hailing a business reality.”
The venture is the latest step toward deploying robo-taxis on American roadways, with several test projects underway and a fully autonomous commercial service deployed in Arizona by Google parent-owned Waymo.