The fleet of bright yellow bikes has been brought to the city by ofo and is the company's first scheme in the north of England.
From today, people will be able to download a free app to locate and unlock a nearby bike, which then charges 50p per half hour to use.
All bikes will be free to hire for the first two weeks, and ofo collects no deposit when registering new users.
Adam Rose, ofo’s operations manager for Sheffield, said: “ofo is set to stir up the way Sheffield residents travel in their city.
"Dockless bikes give people options that can challenge the private car.
"Like countless cities across the UK, Sheffield is marred by air pollution problems, often caused by congestion.
"By bringing ofo to Sheffield we are offering a clean, affordable and practical alternative.
“ofo harnesses the fantastic potential of cutting-edge technology but combines it with the classic utility of the bicycle – something which I’m sure will appeal to the residents of a vibrant and thriving city like Sheffield.”
The scheme has already successfully launched in Cambridge, Oxford, Lodnon and Norwich and it used by 200million people in 20 countries.
Councillor Jack Scott, Sheffield Council's cabinet member for transport and sustainability, said the launch was great news for the city.
He said: "This type of bike sharing is set to change the way we get from A-to-B for the better.
"Not only will ofo provide an affordable and convenient transport option for many, but it will also promote the fantastic health benefits of cycling - reducing air pollution and boosting healthy lifestyles.
"Sheffield Council has been working closely with ofo to ensure that dockless bikes are used responsibly and considerately following their introduction.
"That's why we support ofo's approach of gradually increasing the bikes on Sheffield streets after gauging usage patterns.
"This will help ofo bikes serve the right areas and the right people – enriching and supplementing Sheffield's existing transport options."