The emotional finale was the last of their farewell stage shows in London’s O2 Arena, and the surviving pythons said it would be their final performance together ever.
The men, who are all in their 70s, performed some of their best-loved material including their famous lumberjack song and the dead parrot sketch.
Surprise guests included self-professed Python fans like British theoretical physicist Stephen Hawking and Canada’s own Mike Myers.
On the red carpet before the show, Myers told Reuters how he was influenced by the famous funnymen:
"Python are the mack daddy of comedy. I mean if there was a periodic table of elements of comedy Python would have Pythonium...everybody who does anything, you know, are influenced by Python."
About 15,000 people watched the sold-out show from the O2 Arena, but the festivities were also simulcast to thousands of movie theatres around the world, including a cinema in Calgary.
When weather knocked out their satellite feed, the audience carried on the finale themselves singing Always Look On the Bright Side of Life.