Abstract：Despite the increasing importance of the pig as a large animal model, little is known about porcine amniotic fluid–derived stem (AFS) cells. AFS cells were isolated from 30-day porcine fetuses amniotic fluid. Plasmid containing EGFP gene was transfected into AFS cells by lipofection. The positive AFS cells were abtained. AFS cells were induced to differentiate into cell types representing each embryonic germ layer, including cells of neuronal, adipogenic, osteogenic, myogenic and endothelialand lineages. Expanded AFS cells and differentiated cells were banked or harvested for analysis using reverse transcription–polymerase chain reaction (RT-PCR).Cultured porcine AFS cells widely expressed β-Actin, NogoA, DCX, CyclinD2, CD133, Hes1, Oct4, Desmin, CD-90, Nanog and Sox2. AFS cells were differentiated into astrocyte (GFAP+), oligodendrocyte (GalC+), neuron (NF+、NSE+ and MAP2+), adipocyte (LPL+ and PPARγ-D+), osteoblast (Osteonectin+ and Osteocalcin+), myocyte (myf-6+ and myoD+) and endothelium (CD31+, CD34+, CD144+ and eNOS+). This study shows that porcine AFS cells are broadly multipotent, thus, may be useful in porcine cell transplantation studies potentially leading to the application of this strategy in the setting of disease and injury.
