Bed bug populations are increasing around the world at an alarming rate and have become a major public health concern. The appearance of bed bug populations in areas where Chagas disease is endemic raises questions about the role of these insects in the transmission of Trypanosoma cruzi, the etiological agent of the disease. In a series of laboratory evaluations, bed bug adults and nymphs were experimentally fed with T. cruzi-infected blood to assess the ability of T. cruzi to survive inside the bed bug and throughout the insect's molting process. Live T. cruzi were observed in gut contents of experimentally infected bed bug adults via light microscopy and the identity of the parasite was confirmed via polymerase chain reaction analysis. T. cruzi persisted at least 97-d postinfection in adult bed bugs. Nymphal stage bed bugs that were infected with T. cruzi maintained the parasite after molting, indicating that transstadial passage of T. cruzi in bed bugs took place. This report provides further evidence of acquisition, maintenance, and for the first time, transstadial persistence of T. cruzi in bed bugs.