Barthwell was born and raised in Chicago.[1] Both of her parents are physicians. She received her bachelor's degree from Wellesley College in 2010, where she was the co-president of the improv group Dead Serious.[2] Shortly after graduating she won a scholarship to train in improv and sketch comedy at Second City.[3] She was inspired to apply after seeing Amber Ruffin perform on the Second City Mainstage.[2]
Along with her two brothers and two friends, Barthwell organized the inaugural WakandaCon in Chicago in August 2018, inspired by the release of Black Panther.[1] The event was received positively and drew approximately 2,500 attendees.[6] Barthwell connected the event to afrofuturism and described WakandaCon as "a space for Black people to look past the present and into our future."[6]
Johnathan Appel, Ali Barthwell, Tim Carvell, Liz Hynes, Greg Iwinski, Mark Kramer, Daniel O'Brien, John Oliver, Owen Parsons, Charlie Redd, Joanna Rothkopf, Chrissy Shackelford, Ben Silva, and Seena Vali (2021)
Daniel O'Brien, Owen Parsons, Charlie Redd, Joanna Rothkopf, Seena Vali, Johnathan Appel, Ali Barthwell, Tim Carvell, Liz Hynes, Greg Iwinski, Ryan Ken, Mark Kramer, Sofía Manfredi, John Oliver, Taylor Kay Phillips, and Chrissy Shackelford (2022)
Daniel O'Brien, Owen Parsons, Charlie Redd, Joanna Rothkopf, Seena Vali, Johnathan Appel, Ali Barthwell, Tim Carvell, Liz Hynes, Ryan Ken, Mark Kramer, Sofia Manfredi, John Oliver, Taylor Kay Phillips, and Chrissy Shackelford (2023)
Daniel O'Brien, Owen Parsons, Charlie Redd, Joanna Rothkopf, Seena Vali, Johnathan Appel, Ali Barthwell, Tim Carvell, Liz Hynes, Ryan Ken, Mark Kramer, Sofía Manfredi, John Oliver, Taylor Kay Phillips, and Chrissy Shackelford (2024)