Abstract：Amniotic fluid stem (AFS) cells are the Oct4 positive multipotent stem cells derived from amniotic fluid. These cells possess the robust proliferation, multi-linear differentiation and lowness of immunity and have great promise in different fields of biology and clinical application. In this study, we isolated and characterized the porcine AFS (pAFS) cells devived from different gestational age of porcine fetus, and revealed the correlation between the gestational age of fetus and the proliferation and adhesion rate of pAFS cells. Further more, pAFS cells were able to differentiate into adipocytes when cultured in the special inducing medium. The results showed that pAFS cells derived from the fetus in 40~70 d range obtained the optimal proliferation and adhersion rate. Two porcine AFS cells lines were establishment so far. By FASC and RT-PCR analysis, pAFS cells expressed the following cell surface antigens including CD117, CD44, CD166 and CD90, but negatived for CD34, CD45 and CD54; These cells could be expressed constantly the stem cell special gene markers such as Oct4, Nanog, c-Myc and HLA-abc; The EGPF drived by promoters of Oct4 and Nanog were expressed in pAFS cells when transfected with pOct4-EGFP and pNanog-EGFP; Besides, pAFS cells were successful induced into adipocytes in vitro. This paper presents a method which is useful for the study of AFS cell from domestic animals.
