[Isolation and identification of huamn dental pulp stem cells].
OBJECTIVE
To isolate and identify human dental pulp stem cells from third molars.


METHODS
Dental pulps were dissected and digested by collagenase type I and dispase. The obtained single cell supernatant were harvested and cultured. Characterization of the phenotype of DPSCs was detected by immunohistochemical method and RT-PCR assay. Cell cycle was analyzed by FCM. Differentiation potential of DPSCs was evaluated.


RESULTS
Colony-forming efficiency of cells derived from dental pulp tissue was 2 - 15 clones/10(3) cells plated. DPSCs were found to express many different markers, including vimentin, collagen type I, GFAP, nestin and osteocalcin, while they failed to react with MyoD and DSPP. About 64.1% of the cells were in G0/G1 phases, while only 35.8% in proliferation (S + G2 + M). Grown in an adipogenic cocktail medium for three weeks, some DPSCs expressed fat cell markers of PPARgamma and LPL, and formed oil red O-positive lipid clusters in five weeks. After culture with a myogenic-inductive medium, DPSCs were found to express MyoD, desmin and myosin, markers of myocyte. Long-term cultures of DPSCs grown in differentiation inductive medium demonstrated the capacity to form Von Kossa-positive condensed nodules with high levels of calcium.


CONCLUSIONS
Cells isolated from adult human dental pulp are clonogenic, and have multipotent differentiation potential, satisfying the criteria of postnatal somatic stem cell.
He
Fei
F
Dept. of Conservative Dentistry, Hospital of Stomatology, The Fourth Military Medical University, Xi'an 710032, China.
Tan
Ying-hui
YH
Zhang
Gang
G
D000328:Adult; D002454:Cell Differentiation; D003782:Dental Pulp; D004847:Epithelial Cells; D006801:Humans; D015675:Osteocalcin; D013234:Stem Cells
D015675:Osteocalcin