Optimizing proliferation and characterization of multipotent stem cells from porcine adipose tissue

Porcine mesenchymal stem cells have been isolated previously from bone marrow but not from adipose tissue. In the present study a new cell-culture method, using a low-calcium medium supplemented with N-acetyl-L-cysteine and L-ascorbic acid 2-phosphate (the PM2 medium) was developed to grow pASCs (porcine adipose-tissue-derived stem cells). The pASCs developed using the new medium showed a high growth rate and a high proliferation potential, as measured by a cumulative population doubling level (55) that was significantly higher than those reported for ASCs in the literature. These pASCs lacked gap-junctional intercellular communication and were capable of differentiation into three mesodermal lineages (i.e. adipocytes, osteoblasts and chondrocytes) and an ectodermal lineage (i.e. neural cells). Surprisingly, osteogenic ability, but not adipogenesis, was found to increase dramatically with increasing passages. The high proliferative and differentiation potential of these pASCs should facilitate the development of a large-animal model to study the use of ASCs in regenerative and reparative medicine.