In 1836, the Louisa Railroad Company, the Chesapeake and Ohio Railway's (C&O) oldest predecessor, was chartered. Around 1851, its name was changed to the Virginia Central Railroad. In 1868, special acts of Virginia's and West Virginia's legislatures provided for completion of rail lines from Chesapeake Bay to the Ohio River. Under these acts, the Virginia Central Railroad was renamed the Chesapeake and Ohio Railroad. This company succeeded to the rights, interests and privileges of both the Virginia Central and the Covington and Ohio Railroads.

In 1878, the Chesapeake and Ohio Railroad was renamed the Chesapeake and Ohio Railway (it was reorganized between 1873 and 1878 during receivership).

In 1895, the Baltimore & Ohio Railroad placed the first successful electric locomotive in railroad service. An earlier test of the world's first electric railroad locomotive, the "Page Locomotive," took place on the C&O's Washington Branch in April 1851.

In 1947, the Pere Marquette Railway, principally a Michigan line, was merged into the C&O. Sixteen years later, the C&O acquired stock control of the B&O in February following the ICC's approval on December 31, 1962. In 1966, the ICC authorized the C&O to acquire control of the Chicago South Shore & South Bend Railroad. The next year, the ICC approved control of the Western Maryland Railway by the C&O/B&O. On February 26, 1973, the Chessie System Inc. was formed and Chessie System Railroads was adopted as the new corporate identity for the C&O, B&O and Western Maryland railroads.

Later, the CSX Corporation came into being, resulting from the merger of the Chessie System and Seaboard Coast Line Industries Inc. Three years later, pperation of the Western Maryland Railway was taken over by the B&O, and WM's ownership was assumed by the C&O. In 1987, the B&O was merged into the C&O, and the C&O was merged into CSX Transportation.