ISOLATION AND CULTURE OF PUTATIVE MESENCHYMAL STEM CELLS FROM THE EQUINE AMNIOTIC MEMBRANE

Abstract

Multipotent mesenchymal stromal cells (MSCs) are a promising therapeutic tool for the treatment of equine tendon and other musculoskeletal injuries. While bone marrow is considered the ‘gold standard’ source of these cells, various other tissues contain mesenchymal stem cells with potentially useful features. Equine foetal adnexa, amniotic membranes, chorion, wharton’s jelly, umbilical cord blood (UCB) and amniotic fluid (AF) are potential non-invasive sources of MSCs. We collected amniotic membrane from twenty mares during their foaling for isolation of MSCs and evaluated them for the differences in isolation rates, proliferation capacity and expression of pluripotency markers. The plastic adherent colonies were observed in 80% of the collected samples. The proliferation potential of AM derived MSCs was measured based on their population doubling time (PDT). Amnion derived MSCs expressed CD29, CD44, CD73, CD90 and CD105 and were negative for haematopoietic and leukocytic markers (CD14, CD34 and CD45). These MSCs also expressed the Oct-4, Sox-2 and Nanog, which are the pluripotency markers. The present study results indicated that amniotic membrane derived MSCs may be a good source of putative stem cells, making them potentially useful for veterinary regenerative medicine and cell-based therapy.