Vojnosanitetski pregled 2014 Volume 71, Issue 8, Pages: 735-741
https://doi.org/10.2298/VSP1408735D
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Characterization of deciduous teeth stem cells isolated from crown dental pulp
Debeljak-Martačić Jasmina (Institute for Medical Research, Belgrade)
Francuski Jelena (Faculty of Veterinary Medicine, Department of Pathophysiology, Belgrade)
Lužajić Tijana (Faculty of Veterinary Medicine, Department of Pathophysiology, Belgrade)
Vuković Nemanja (University of Business Academy, Faculty of Stomatology, Pančevo)
Mojsilović Slavko (Institute for Medical Research, Belgrade)
Drndarević Neda (University of Business Academy, Faculty of Stomatology, Pančevo)
Petakov Marijana (University of Business Academy, Faculty of Stomatology, Pančevo)
Glibetić Marija (Institute for Medical Research, Belgrade)
Marković Danica (Faculty of Veterinary Medicine, Department of Pathophysiology, Belgrade)
Radovanović Anita (Faculty of Veterinary Medicine, Department of Pathophysiology, Belgrade)
Todorović Vera (University of Business Academy, Faculty of Stomatology, Pančevo)
Kovačević-Filipović Milica (Faculty of Veterinary Medicine, Department of Pathophysiology, Belgrade)
Background/Aim. The last decade has been profoundly marked by persistent
attempts to use ex vivo expanded and manipulated mesenchymal stem cells
(MSCs), as a tool in different types of regenerative therapy. In the present
study we described immunophenotype and the proliferative and differentiation
potential of cells isolated from pulp remnants of exfoliated deciduous teeth
in the final phase of root resorption. Methods. The initial adherent cell
population from five donors was obtained by the outgrowth method. Colony
forming unit-fibroblast (CFU-F) assay was performed in passage one. Cell
expansion was performed until passage three and all tests were done until
passage eight. Cells were labeled for early mesenchymal stem cells markers
and analysis have been done using flow cytometry. The proliferative potential
was assessed by cell counting in defined time points and population doubling
time was calculated. Commercial media were used to induce osteoblastic,
chondrogenic and adipogenic differentiation. Cytology and histology methods
were used for analysis of differentiated cell morphology and extracellular
matrix characteristics. Results. According to immunophenotype analyses all
undifferentiated cells were positive for the mesenchymal stem cell markers:
CD29 and CD73. Some cells expressed CD146 and CD106. The hematopoietic cell
marker, CD34, was not detected. In passage one, incidence of CFU-F was 4.7 ±
0.5/100. Population doubling time did not change significantly during cell
subcultivation and was in average 25 h. After induction of differentiation,
the multicolony derived cell population had a tri-lineage differentiation
potential, since mineralized matrix, cartilage-like tissue and adipocytes
were successfully formed after three weeks of incubation. Conclusion.
Altogether, these data suggest that remnants of deciduous teeth dental pulp
contained cell populations with mesenchymal stem cell-like features, with a
high proliferation and trilineage differentiation potential and that these
cultures are suitable for further in vitro evaluation of cell based
therapies.
Keywords: dental pulp, stem cells, tooth, deciduous, child, preschool, cell differentation, adipogenesis, chondrogenesis, osteogenesis
Projekat Ministarstva nauke Republike Srbije, br. 175061