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Canine mesenchymal stem cells (MSCs) can be defined with self renew potential and specific differentiation capacity. Amiotic membrane represent an important source of MSCs, which can be harvested by minimally invasive methods. The aim of our study was to evaluate the growth characteristics of canine amniotic membrane derived mesenchymal stem cells. The placenta samples were collected after cesarean section from healthy mixed breed dogs. MSCs isolation was performed using enzymatic method. Isolated cells were cultured in propagation medium: Dulbecco’s Modified Eagle’s Medium/F12 (DMEM/F12, Gibco) supplemented with 10% fetal bovine serum (FBS, Gibco) and 1% antibiotic-antimycotic (Sigma-Aldrich). The medium was changed after 4 days. The cell doubling number, cell proliferation capacity, cell doubling time, daily duplication rate and clonogenic efficacy were evaluated. Our study demonstrate the self renew potential of canine amniotic membrane derived mesenchymal stem cells, and can represent a potential source of stem cells for canine regenerative medicine.