The 23-year-old Wilson was told by doctors Monday that he risked more serious problems if he kept playing.
He underwent fusion surgery to repair vertebrae and a herniated disk in his neck in January. Then at practice last Tuesday, Wilson caught a pass and ran with his head down into the back of an offensive lineman, a hit that caused numbness in his hands and lower extremities.
He missed the final 11 games last season after being diagnosed with spinal stenosis, a narrowing of the spinal cord.
''I don't want anybody to feel sorry for me, or pity me,'' he said in a statement released by the team. ''I lived my dream. A lot of people only get to dream their dream. I lived that dream. Now I have a chance to dream another dream and live that, too.''
Wilson was drafted in the first round, 32nd overall, out of Virginia Tech in 2012. As a rookie, he rushed for 358 yards and scored four touchdowns while making his mark as a kick returner. Wilson led the NFL with 1,533 kickoff return yards, a team record.
But he got off to a slow start last season before the injury.
Wilson's condition is not expected to require additional surgery or therapy.
Wilson met Monday morning with Dr. Russell Warren, the team's physician, and Dr. Frank Cammisa, the chief of spine service at New York's Hospital for Special Surgery. Cammisa performed the spinal fusion surgery on Wilson.
They told him he needed to stop playing football.
Wilson then returned to the Giants' training facility and met with team President John Mara, general manager Jerry Reese and coach Tom Coughlin.
Coughlin was prepared for the worst before getting the official medical reports about Wilson.
''We had pretty much braced ourselves for not good news, although trying to be optimistic and not trying to go ahead and make some kind of a statement prior to getting that information,'' he said on a conference call. ''We were certainly excited thinking that everything was going to be OK with David. But in the back of your mind: How are you going to be affected as a team? It would be irresponsible not to give some thought to that.''
Wilson had only 146 yards on 44 carries and one touchdown in five games last season before the injury. He struggled with holding onto the ball, losing three fumbles.
Against New Orleans his rookie season, Wilson set a team record for all-purpose yards with 327. He had 227 yards on kickoff returns and added 100 rushing.
The Giants will now move on without Wilson. In the preseason opener against Buffalo on Sunday, rookie Andre Williams led a power running game. He had 48 yards on seven carries and a TD. Veteran Peyton Hillis added 36 yards on seven attempts, and newcomer Rashad Jennings gained 23 yards on seven carries and caught three passes for 20 yards.
''I'm encouraged, period,'' Coughlin said. ''I thought eventually we got to where we ran the ball pretty well. That might not have been their (starters) in there, but still we did have some consistency with runs there.''