Comparison of osteo/odontogenic differentiation of human adult dental pulp stem cells and stem cells from apical papilla in the presence of platelet lysate.
BACKGROUND
Human dental pulp cells (DPSCs) and stem cells from apical papilla have been used for the repair of damaged tooth tissues. Human platelet lysate (PL) has been suggested as a substitute for fetal bovine serum (FBS) for large scale expansion of dental stem cells. However, biological effects and optimal concentrations of PL for proliferation and differentiation of human dental stem cells remain to be elucidated.


METHODS
DPSCs and SCAP cells were isolated from impacted third molars of young healthy donors, at the stage of root development and identified by markers using flow cytometry. For comparison the cells were cultured in media containing PL (1%, 5% and 10%) and FBS, with subsequent induction for osteogenic/odontogenic differentiation. The cultures were analyzed for; morphology, growth characteristics, mineralization potential (Alizarin Red method) and differentiation markers using ELISA and real time -polymerase chain reaction (qPCR).


RESULTS
The proliferation rates of DPSCs and SCAP significantly increased when cells were treated with 5% PL (7X doubling time) as compared to FBS. 5% PL also enhanced mineralized differentiation of DPSCs and SCAP, as indicated by the measurement of alkaline phosphatase activity, osteocalcin and osteopontin, calcium deposition and q-PCR.


CONCLUSIONS
Our findings suggest that using 5% platelet lysate, proliferation and osteo/odontogenesis of DPSCs and SCAP for a short period of time (15 days), was significantly improved. This may imply its use as an optimum concentration for expansion of dental stem cells in bone regeneration.
Cell therapy center, Faculty of Medicine, University of Jordan, Amman, Jordan. Electronic address: duaa_abuarqoub@yahoo.com.
Duaa
D
Abuarqoub
Cell therapy center, Faculty of Medicine, University of Jordan, Amman, Jordan.
Abdalla
A
Awidi
Department of applied Biology, Jordan University of Science and Technology, Irbid, Jordan.
Nizar
N
Abuharfeil
D000293:Adolescent; D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology* / Q000378:metabolism; D001792:Blood Platelets / Q000166:cytology*; D002450:Cell Communication / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D003782:Dental Pulp / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D008964:Molar, Third / Q000166:cytology; D009805:Odontogenesis / Q000502:physiology; D015675:Osteocalcin / Q000378:metabolism; D010012:Osteogenesis / Q000502:physiology; D055815:Young Adult
D015675:Osteocalcin
DPSC; Dental stem cells; Platelet lysate; SCAP