The Soccer City stadium, close to the township of Soweto, was hit by major traffic jams for a second time on Thursday when South Africa beat Colombia in a pre-World Cup friendly.
Many fans abandoned their cars on the side of the road and walked to the stadium.
They still missed part of the game, repeating problems seen last weekend when the South African cup final coincided with a major rugby game at another stadium nearby.
After those incidents, officials blamed the coincidence of two games, World Cup roadworks and a transport strike, but on Friday both they and spectators said the fault was with fans who refused to use public buses and the park-and-ride system.
Sibongile Mazibuko, the top Johannesburg city official for the World Cup, said the plan for Soccer City, newly inaugurated as the biggest stadium in Africa, and Ellis Park, another venue for the tournament, was based on using public transport.
"It can't be the convenience of using your own car at the expense of inconveniencing the whole tournament. We won't let it happen, we will tow people's cars if they attempt to come to the stadium with them," she told Reuters.
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