The decision comes a day after Spurs were trashed 6-1 at Newcastle – a performance chairman Daniel Levy has described as “wholly unacceptable”.
Ryan Mason, who worked under Stellini and took charge when Jose Mourinho was sacked in 2021, succeeds him.
Sunday’s defeat at Newcastle, where they trailed 5-0 after 21 minutes, was a significant blow to Spurs’ hopes of securing Champions League qualification.
“It was devastating to see,” added Levy.
“We can look at many reasons why it happened and while myself, the board, the coaches and players must all take collective responsibility, ultimately the responsibility is mine.”
Tottenham are fifth in the Premier League table, six points behind both Newcastle in third and Manchester United in fourth.
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Both teams have played fewer games than Spurs, who host Erik ten Hag’s side on Thursday.
Spurs won one, drew one and lost two games after appointing Stellini, who acted as Conte’s assistant during his time as manager.
Stellini also led the side when Conte was missing from the touchline after a gall bladder operation in February, with Spurs winning three of their four Premier League games in Conte’s absence, but also losing to Championship side Sheffield United in the FA Cup.
“Cristian stepped in at a difficult point in our season and I want to thank him for the professional manner in which he and his coaching staff have conducted themselves during such a challenging time,” said Levy.
Former Spurs and England midfielder Mason, 31, returns to the helm for the final six games of the season.
“Ryan knows the club and the players well,” added Levy.