Abstract
The dental pulp represents an easily acces-sible source of adult dental pulp stem cells (DPSCs). The preferred approach to DPSC isolation is enzy-matic digestion.
However, the duration of the enzymat-ic activity is crucial. The purpose of this study was to isolate the DPSC populations using this method, characterize their biological properties and prolifer-ation capacity, and to determine their ability to dif-ferentiate into mature cells.
Before enzymatic diges-tion using 0.05% trypsin, we used the homogenization method in order to obtain a fine homogenate from the solid pulp tissue. The stem cells were cultivated in modified cultivation medium for mesenchymal adult progenitor cells containing 2% foetal bovine serum, growth factors and insulin-transferrin-sele-nium supplement.
We were successfully able to iso-late 10 populations of DPSCs. The vitality of DPSCs did not drop below 90 %.
However, the DPSCs showed a significant decrease in the relative telomere length number with increasing passaging (P < 0.05). Isolated DPSCs highly expressed the CD markers: CD29, CD44, CD90, CD13, CD73 and CD166.
In con-trast, CD markers CD31, CD106, CD34 and CD45 were negative or low positive. We confirmed the high osteogenic and chondrogenic potential of the isolated stem cells.
Isolated DPSCs did not show signs of cell degeneration or spontaneous differentiation during the entire cultivation. In addition, we were able to shorten the enzyme activity duration, and we were the first to demonstrate trypsin as the enzyme used for the enzymatic digestion method with the viability over 90 % of isolated DPSCs using this method.