In pig-to-human xenotransplantation, zoonotic infections have been an important barrier. The risk of zoonosis has been emphasized in xenotransplantation after finding that porcine endogenous retroviruses (PERVs) can infect human cells in vitro. Until now, transmissions of PERVs from PK15 cells have been studied in vitro and in vivo, but transmission of PERVs originating from miniature pigs have not been extensively reported. Peripheral blood mononuclear cells from miniature swine showed PERV transmission to human cells. In contrast, specific pathogen-free (SPF) pig islet cells showed no PERV transmission when co-incubated with 293T cells. To evaluate the risk of zoonosis with our experimental mini pigs, we tested the infectivity of PERVs from NIH-miniature pig primary ear cells for human 293T cells. As a result, all subgroups of infectious PERV virion (PERV-A, -B, and -C) were detected in the primary cell culture media. Unlike PERV-C, PERV-A and -B infected human 293T cells. Interestingly, only proviral PERV-A replicated in 293T cells to produce virions after infection. Our results suggested that a prevention study of PERV xenotransmission from experimental miniature pigs should concentrate on PERV-A control.