The Chesapeake & Ohio Railway (C&O, CO) was a Class I railroad formed in 1869 in Virginia from several smaller Virginia railroads begun in the 19th century. Led by industrialist Collis P. Huntington, it reached from Virginia's capital city of Richmond to the Ohio River by 1873, where the railroad town (and later city) of Huntington, West Virginia, was named for him.

By the early 1960s the C&O was headquartered in Cleveland, Ohio. In 1972, under the leadership of Cyrus Eaton, it became part of the Chessie System, along with the Baltimore and Ohio and Western Maryland Railway. The Chessie System was later combined with the Seaboard Coast Line and Louisville and Nashville, both the primary components of the Family Lines System, to become a key portion of CSX Transportation (CSXT) in the 1980s.