Colosimo has had some success on film, mainly in Australia. He made his film debut in the coming-of-age story Moving Out in 1983 and featured in 1984's Street Hero.

Despite several movie roles and guest roles in A Country Practice (1994), Good Guys Bad Guys (1997) and Stingers (2002), it was not until his performance as Joe Sabatini in Something in the Air in 2001 and 2002 that resulted in wider exposure. In 2003, he starred in the telemovie, After the Deluge. In 2003 and 2004, he played Dr. Rex Mariani in The Secret Life of Us. Vince also starred as himself in episode 6 of the hit ABC series We Can Be Heroes. He is chosen as the actor to play Phil Olivetti in the fictional mini-series within the show.

Other credits include the cult movie Chopper (2000), in which he played Melbourne drug dealer Neville Bartos opposite Eric Bana, The Wog Boy (2000) Lantana (2001), Walking on Water (2002), The Nugget (2002), Take Away (2004) and Opal Dream (2006). In 2008, he starred alongside Leonardo DiCaprio in the 2008 American film Body of Lies. In 2010, Colosimo starred in Wog Boy 2: Kings of Mykonos (the sequel to The Wog Boy). Colosimo also appeared in the vampire film Daybreakers, starring alongside Sam Neill, Willem Dafoe and Ethan Hawke.

Colosimo was born in Melbourne, one of four children of Italian-born parents from Calabria. He grew up in the inner city suburb of Carlton North. He has a daughter, Lucia (2002), with his former wife, Actress Jane Hall. Colosimo and Hall worked together on A Country Practice in 1994. He lived in Westgarth, a suburb of Melbourne, until late 2015. The mother of his second child is Australian Actress Diana Glenn with whom he had a son, Massimo, in April 2014. The couple separated 2 months after the birth of Massimo.

From 2005 until late 2007, Colosimo had guest roles in Blue Heelers (2005), MDA (2005), Two Twisted (2006) and City Homicide (2007). His American credits include popular shows such as The Practice (2004) and Without a Trace (2003). In 2008, Colosimo had a busy year. He portrayed Melbourne gangland figure Alphonse Gangitano in the Channel 9 series Underbelly (in a guest role), played in Channel Ten's telemovie Emerald Falls and Channel Nine's Scorched. He also appeared in Top Gear Australia's "Star in a bog-standard car" section in the first episode.

He also sampled his voice on Snitch by Vanessa Amorosi in on her 2009 album Hazardous (album)

He was selected as one of the entrants to the Who's Who in Australia 2011 edition.

In 2012 he and his ancestors featured in the SBS show, Who Do You Think You Are?

He announced in late 2013 that he would be reprising his role of Alphonse Gangitano in the sequel/spin-off series Fat Tony & Co. which will not be placed under the Underbelly franchise (due to financing reasons) and will focus on the rise and fall of Tony Mokbel, who also featured in the original series, played by Robert Mammone.

In September 2016, Colosimo was charged by police after he was found in possession of Methamphetamine in Melbourne's north. In January 2017, he appeared in court where he had pleaded guilty and was given a fine of $1000 without conviction and was put under a good behaviour bond for a year.