The train type was introduced into service on 31 March 1991 following the introduction of Hokuso Kodan Line through services, and to replace the 5000 series trainsets that had been used on the Asakusa Line since its inception in 1960.

Passenger accommodation consisted of longitudinal bucket seating throughout.

[7] After the speed limit of the Keikyu Line was raised to allow for 120-kilometre-per-hour (75 mph) operation between Shinagawa and Yokohama stations in 1995, a 5300 series set (5327) entered service in 1998 with uprated traction motors (to 180 kW or 240 hp[1]: 132 ) to support such operation.

[6] Following the introduction of new Toei 5500 series trainsets,[8] withdrawals commenced on 27 July 2018, with set 5301 being the first to be withdrawn; it was scrapped on 1 August of that year.

[9] Set 5320, the last trainset in operation,[7] made its final trip in revenue service in February 2023.