The 38-year-old Stepanek won five ATP singles titles and reached the Wimbledon quarter-finals in 2006 when he reached a career-high eighth in the world rankings.
The following year a back injury threatened his career but Stepanek battled back and won two grand slam double titles with India's Leander Paes, the Australian Open in 2012 and the U.S. Open a year later.
Alongside Tomas Berdych, Stepanek was also part of the Davis Cup teams triumphed in 2012 and 2013.
Stepanek had hoped to make another comeback after undergoing back surgery but that dream was never realised.
"I had to be realistic. I fought till the last breath but it is time to move on," Stepanek was quotes as saying on www.idnes.cz website.
(Reporting by Jan Lopatka, editing by Pritha Sarkar)
