Isolation of multipotent stem cells from adult rat periodontal ligament by neurosphere-forming culture system.
Adult multipotent stem cells have been isolated from various non-neural tissues. Here, we report the isolation of multipotent stem cells from rat periodontal ligament (PDL) using neurosphere-forming culture system. Enzymatically dissociated PDL cells were cultured in serum-free basal medium containing EGF, bFGF, and LIF. Free-floating spheres expressing nestin, GFAP, and vimentin were formed by 7 days of the culture. In addition, spheres expressed mRNA of neural crest-associated transcription factors Twist, Slug, Sox2, and Sox9. PDL-derived spheres differentiated into multinucleated myotubes, NFM-positive neuron-like cells, GFAP-positive astrocyte-like cells and CNPase-positive oligodendrocyte-like cells. Methylcellulose colony-forming assay revealed that a single PDL cell could form a sphere at a frequency of approximately 0.01% of total cells. These data indicate that PDL-derived spheres contained multipotent adult stem cells capable of differentiating into both neural and mesodermal progeny. This is the first report of the isolation of PDL-derived stem cells with primitive neural crest stem cell features.
Techawattanawisal
Wanida
W
Department of Cellular Physiological Chemistry, Graduate School, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8549, Japan.
Nakahama
Kenichi
K
Komaki
Motohiro
M
Abe
Mayumi
M
Takagi
Yuzo
Y
Morita
Ikuo
I
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D008297:Male; D039902:Multipotent Stem Cells / Q000166:cytology*; D009474:Neurons / Q000166:cytology*; D010513:Periodontal Ligament / Q000166:cytology*; D051381:Rats; D017208:Rats, Wistar; D023822:Tissue Engineering / Q000379:methods*