Ninety-six people - mostly Liverpool fans - were crushed to death at the Sheffield Wednesday stadium in 1989 after too many fans were allowed into the stadium during an FA Cup tie against Nottingham Forest.
The Roman epic, which was directed by Ridley Scott, has been released on DVD along with an additional documentary on the links between sport and violence.
However, because of an error at the US library which supplied pictures to its producers, an image from the 1989 tragedy was included in the documentary.
The error was not spotted until a support group for relatives of the victims got in touch with its UK distributor, Columbia TriStar.
Ninety-six people - mostly Liverpool fans - were crushed to death at the Sheffield Wednesday stadium in 1989 after too many fans were allowed into the stadium during an FA Cup tie against Nottingham Forest.
The still has now been removed from the documentary, which was produced separately from the Oscar-nominated film.
'Sincere apologies'
A Columbia TriStar spokesman said the company, together with studios DreamWorks and Universal, "deeply regretted" the error and offered their "sincere apologies".
"We are sincerely sorry for any additional pain this may have caused," he said.
"Now we are aware of this we have taken steps to remove this photograph from all future copies of the Gladiator DVD."
Gladiator was the biggest-selling DVD of 2000, selling more than half a million copies. It helped push DVD sales up to 16.6 million over the year - four times the amount sold in 1999.
The film itself has been nominated for 15 Bafta awards, and is widely tipped to feature highly in Tuesday's Academy Award nominations.