After captaining Glasgow City to a 14th consecutive SWPL title the former Falkirk Girls player, 39, will join the club’s first team coaching staff as assistant coach.
Commenting on her decision, Ross said: “It hasn’t been an easy decision to call time on my playing career but I feel now is the right time.
“I have spent 14 happy and successful years at Glasgow City and have been privileged to have played with and captained so many amazing people and footballers.
"This club means so much more to me than just football, it has become my extended family.
"I have so many special memories and have made lifelong friends, it’s a journey I wouldn’t change for the world.
“I am excited to embark on a new chapter of my career and am delighted it is with Glasgow City, a place that I call home.”
Along with the league titles, Ross won six Scottish Women’s Premier League Cups and seven Scottish Cups and retires as the most decorated Scottish player ever.
Ross joined the club in the summer of 2007 and made her Glasgow City debut in the opening league game of the 2007-08 away to Aberdeen.
By the end of the season, she would be a league champion, a title that would remain for every single season at City.
At international level, Ross won 133 Scotland caps and scored nine goals before retiring from international football in 2017.