i Sebastiani was founded on June 4th, 1990 as a splinter organization of the Society for Creative Anachronism (SCA). Founder Jeff Hatalsky's stated objective was to drive the group towards greater authenticity. In September 2001, i Sebastiani attended the Austin Commedia dell'Arte Festival in Austin Texas, where they were met with acclaim. From 1991 through September 2001, the Troupe had performed each Commedia dell'Arte scenario only once, but as demand for Commedia built, i Sebastiani began to perform multiple times with each scenario.

    In October 1996, Jeff Hatalsky left the troupe, and it began a period of attempted democracy. This mode of organization imposed natural limitations, and through reform Alex Newman stepped beyond his role as coordinator to become the troupe's new manager. In October 2002, the troupe expanded beyond SCA events and began to perform for the general public.

    i Sebastiani's Florida debut came at the Miami Improv Festival in 2006. Octavio Roca of the Miami New Times reported on the troupe's involvement: "Their masks hark to the Renaissance, and their stock characters, including Arlecchino and Pantalone, are archetypes. But their plots come from the audience and — within the rough scenario that begins it all — their humor is of the streets."

    i Sebastiani frequently performs to packed crowds at the Charles River Museum of Industry in Waltham, MA. A portion of the proceeds from recent shows have gone towards the repair of flooding damage. The damage was incurred by the Museum during historic rainfall in spring of 2010, when the Charles River reached a century high-water level.