Acta Med. 2010, 53: 79-84

Osteogenic Differentiation of Human Dental Pulp-derived Stem Cells under Various Ex-vivo Culture Conditions

Jana Karbanováa,b, Tomáš Soukupa, Jakub Suchánekc, Jaroslav Mokrýa

aCharles University in Prague, Faculty of Medicine and University Hospital Hradec Králové, Department of Histology and Embryology, Hradec Králové, Czech Republic
bTissue Engineering Laboratories, BIOTEC, Technische Universität Dresden, Germany
cCharles University in Prague, Faculty of Medicine and University Hospital Hradec Králové, Department of Dentistry, Hradec Králové, Czech Republic

Received March 5, 2010
Accepted March 19, 2010

Dental pulp stem cells (DPSCs) can be easily isolated and cultured in low-serum containing medium supplemented with growth factors PDGF-BB and EGF while exhibiting multipotency and immature phenotypic characteristics. In the present study, we investigated their potential to differentiate towards osteogenic lineages using various culture conditions in order to optimize their therapeutic use. DPSCs were cultured either as a cell monolayer or as three-dimensional (3D) micro-mass structures. Monolayers preincubated with bFGF and valproic acid for one week prior their differentiation were cultured in serum containing standard osteodifferentiation medium for four weeks, which resulted in multilayered nodule formation. Micro-mass structures were cultured for same period either in serum containing medium or under serum-free conditions supplemented with TGF-β3 with or without BMP-2. Histochemically, we detected massive collagen I and weak calcium phosphate depositions in multilayered nodules. When culture 3D-aggregates in either standard osteodifferentiation medium or serum-free medium containing TGF-β3, only small amount of collagen I fibres was observed and almost no deposits of calcium phosphate were detected. In contrast, in presence of both TGF-β3 and BMP-2 in the serum- free medium a significant amount of collagen I fibers/bundles and calcification were detected, which is in line with osteogenic effect of BMP-2. Thus, our data indicate that certain environmental cues can enhance differentiation process of DPSCs into osteogenic lineage, which suggest their possible utilization in tissue engineering.


This work was supported by Grant Agency of the Czech Republic (GACR 304/07/P307) and Ministry of Education, Youth and Sports of the Czech Republic (MSM 0021620820).


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