Di Stefano was taken to the intensive care unit at Madrid's Gregorio Maranon Hospital on Saturday after suffering a heart attack and was reported to be in a critical condition.
And on Monday, Real confirmed that Di Stefano had passed away, provoking an outpouring of tributes from around the football world.
Widely regarded as one of the greatest players to ever grace the game, Di Stefano leaves behind a legacy that will never be forgotten.
Di Stefano made his name during an 11-year spell at Real, whom he eventually joined following a dispute between the Spanish capital club and fierce rivals Barcelona over his signature from Colombian outfit Millonarios.
Before that, the Argentina-born forward began his career at River Plate, but it was at the Bernabeu where he really shot to stardom, with an incredible 308 goals in 396 matches making him the club's second-highest scorer behind Raul.
After joining in 1953, 'The Blond Arrow' played an integral role in guiding the Spanish giants to five consecutive European Cups between 1956 and 1960.
Di Stefano scored in every final, including a hat-trick in the famous 7-3 thumping of Eintracht Frankfurt in the last of those.
As well as achieving European glory, Di Stefano helped Real dominate the domestic scene, winning eight La Liga titles in his time at the club, as well as the Copa del Rey in 1962.
Di Stefano - capped for Argentina before acquiring Spanish citizenship and playing for his adopted national side - won the Ballon d'Or twice, in 1957 and 1959, and after leaving Real had a short spell at Espanyol, where he finished his career in 1966.
A move into management followed, with Di Stefano taking the reins at a number of clubs, including two spells at Real.
He was unable to deliver the success as a coach that he had as a player, but his legendary status at the Bernabeu was confirmed in 2000 when new president Florentino Perez announced that Di Stefano was to become honorary president.
The conclusion of the 2013-14 season saw Real clinch their 10th European Cup triumph - La Decima - but it was the feats of Di Stefano in the 1950s and 60s that began the club's obsession with the iconic trophy.
And the achievements of Di Stefano, Ferenc Puskas and co will surely never be matched.
