The first-team duo have made the interim manager's 23-man list for this month's World Cup qualifiers against Malta and Slovenia.
The Three Lions will gather at St George's Park on Tuesday, ahead of Saturday's clash with Malta at Wembley. A trip to Ljubljana follows for the game against Slovakia on Tuesday.
Stones and Sterling have both impressed for City this season, under the leadership of Pep Guardiola.
On-loan goalkeeper Joe Hart has also been selected.
Southgate said: “We've tried to keep some continuity from last month because my view would be that there are a lot of things that have been going right.
"It's obviously been a difficult week in a number of ways, however we are in a position now where it's important to get the squad named.
"I'm looking forward now to working with the players because it's important that we build on the performance and the result against Slovakia."
Fraser Forster (Southampton), Joe Hart (Torino, loan from Manchester City), Tom Heaton (Burnley), Ryan Bertrand (Southampton), Gary Cahill (Chelsea), Phil Jagielka (Everton), Glen Johnson (Stoke City), Danny Rose (Tottenham Hotspur), Chris Smalling (Manchester United), John Stones (Manchester City), Kyle Walker (Tottenham Hotspur), Eric Dier (Tottenham Hotspur), Jordan Henderson (Liverpool), Alex Oxlade-Chamberlain (Arsenal), Michail Antonio (West Ham United), Jesse Lingard (Manchester United), Raheem Sterling (Manchester City), Theo Walcott (Arsenal), Dele Alli (Tottenham Hotspur), Wayne Rooney (Manchester United), Marcus Rashford (Manchester United), Daniel Sturridge (Liverpool), Jamie Vardy (Leicester City)