The Boston and Worcester Railroad began construction of the Milford Branch in early 1847, heading south from their mainline at Framingham, and opened from Framingham to Holliston by September 1847.

By 1958, the line had only one train per day in each direction, running as a shuttle to/from Framingham during the rush hour only and serving Holliston and Milford stations only.

[8] Freight service still continued under the B&A, but the line was by this point an unimportant branchline.

Eventually, as with the rest of the former B&A, the Milford Branch came under the control of Penn Central in their 1968 creation.

[11][12][13] Starting in 2017, portions of the rail line began being converted into the Upper Charles Rail Trail, which as of 2024 has been completed from Whitney Street in Sherborn south to Downtown Milford.