The 32-year-old, who impressed for the Cheetahs during the Currie Cup this season following his return home from a spell with French club side Toulon, has taken the decision to retire from the game with immediate effect.
The Cheetahs are one of four Currie Cup sides to take on the Lions next summer but Rossouw will not be a part of the June 6 clash after he was advised by medical experts to call time on his career.
"I spoke to a number of specialists and decided to call it a day while I am still fine," said Rossouw.
"I was still enjoying playing the game, but I'm looking forward to taking my career into a new direction."
Rossouw had been expected to feature prominently in next year's Super 14 but a number of recurring concussions has meant he will now look at coaching or business options instead.
The former Western Province and Stormers pivot had hoped to follow in the footsteps of his elder brother Pieter, who faced the Lions on two occasions back in 1997.
Pieter started both the second and third Tests against Ian McGeechan's men 11 years ago, scoring a try in the Boks' 35-16 win in Johannesburg. The Lions' second test win in Durban was Rossouw's first of 21 matches in Springbok colours.