Staff and volunteers at the Disability Community in Possilpark, Glasgow, which appeared on Channel 4's Secret Millionaire in 2008, say they have reached crisis point.
The 30-year-old organisation runs social clubs and dozens of classes, as well as after-school care for people with a range of physical and learning disabilities.
In the show, property tycoon Nick Leslau spent time with service users and staff and donated £225,000, which was used to upgrade the building.
But the organisation now needs to raise £50,000 to secure its future.
Madonna Maynes, adult and youth coordinator, said: "It would leave a hole in the community but we don't think we can operate any longer. The staff know they won't be paid at the end of the month - there's no money there.
"We desperately need funding to keep going."