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The 31-year-old said it was "time to move on" as she made the announcement in an Instagram post.
She wrote: "Thank you cycling for everything you've given me - including a husband and our growing family!
"Having people say I have inspired women and girls to get active and get on a bike means the world to me.
"Thanks to Team GB, British Cycling and all the partners who have supported my journey. A special thanks to every team mate I have had over the years and of course to my family for being the best support unit I could ever have wished for."
Dame Laura added: "It's now time to move on but stay following for the next chapter."
She won five Olympic golds and seven world titles during her career on the track. She is married to former cyclist Jason Kenny, who is the most successful British Olympian.
The couple have two sons - born in 2017 and 2023.
Dame Laura suffered a miscarriage in November 2021 before being rushed into surgery two months later with an ectopic pregnancy.
Her retirement comes less than two weeks after British Cycling's performance director Stephen Park said there was only a "slim chance" of her competing at the Paris Olympics this summer.
She had been hoping to make a fourth Olympics appearance but had not returned to Team GB training after giving birth last July.
She first won gold in the women's omnium and team pursuit at the London Olympics in 2012 and repeated this achievement in Rio in 2016.
At the Tokyo Olympics, which was delayed to 2021 because of the pandemic, Kenny won team pursuit silver as well as gold with Katie Archibald in the women's madison - a new event at that year's Games.
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As well as being the most decorated female Olympian, she is the first British woman to win gold medals in three consecutive Olympics.
Her last major honour was winning gold in the scratch race at the 2022 Commonwealth Games in Birmingham.