Abstract: The study was designed to establish a method for the efficient gene transfection in vitro and rapid screening of amniotic fluid-derived stem cells (AFSC), while maintaining their pluripotency. pCMV-DsRed was used as the reporter plasmid to transfect amniotic fluid-derived stem cells by liposome. 36 h later, transient transfection efficiency was 69.17% by flow cytometry. The purified transgenic sheep AFSC monoclones were obtained through the selection of G418 at the concentration of 6 mg/mL after the treatment of 5 h. The transgenic sheep AFSC was positive for Oct-4, SSEA-1 by RT-PCR. AFSC could also formed embryoid bodies by in vitro suspension culture. The results implied a possibility that the transgenic sheep AFSC could be used to track down in vivo differentiation path and the definitive host.

Key words: sheep; amniotic fluid stem cells; reporter plasmid; gene transfection; efficient transfection; rapid screening