Derivation of Presumptive Stem Cells from the Bovine Amnion

Amnion is a noncontroversial, inexhaustible source that can be harvested noninvasively at low cost; moreover, presumptive stem cells are not acutely rejected when transplanted. As in human medicine, stem cells isolated from animal term placenta are promising candidates for disease treatment, specifically for their plasticity and their reduced immunogenicity and high anti-inflammatory potential. The aim of this work was to obtain, for the first time, presumptive stem cells from the epithelial (AECs) and mesenchymal (AMCs) portions of the bovine amniotic membrane to be used for clinical applications, in particular for the treatment of mastitis. AECs and AMCs were isolated following the method used by Lange Consiglio et al. (2010) in horse. Our preliminary results show that cells from epithelial portion displayed a polygonal morphology whereas those from mesenchymal site were fibroblastic-like. Both cell lines were able to adhere to the plastic dishes and resulted positive for MSC-markers (CD44, CD29, CD105, CD73 and CD166) and negative for the hematopoietic marker (CD34) from passage 1 (P1) to P3. Obtained cells showed a pluripotent profile by expressing c-Myc, Oct-4 but not Nanog at every passage studied. AECs and AMCs MHC-I at every passage whereas only AMCs did not express MHC-II at the first passage. Further studies, including pre-clinical and a deeper evaluation of immunological properties, need to be performed for the in vivo applications in order to better understand the role of these cells in cellular therapy.