Isolation of multipotent stem cells from adult rat periodontal ligament by neurosphere-forming culture system

摘要: Adult multipotent stem cells have been isolated from various non-neural tissues. Here, we report the isolation of rat periodontal ligament (PDL) using neurosphere-forming culture system. Enzymatically dissociated PDL were cultured in serum-free basal medium containing EGF, bFGF, and LIF. Free-floating spheres expressing nestin, GFAP, vimentin formed by 7 days culture. In addition, expressed mRNA neural crest-associated transcription factors Twist, Slug, Sox2, Sox9. PDL-derived differentiated into multinucleated myotubes, NFM-positive neuron-like cells, GFAP-positive astrocyte-like CNPase-positive oligodendrocyte-like cells. Methylcellulose colony-forming assay revealed that a single cell could form sphere at frequency approximately 0.01% total These data indicate contained adult capable differentiating both mesodermal progeny. This is first with primitive crest features.