Evaluation of immunophenotyping, proliferation and osteogenic differentiation potential of SSEA-4 positive stem cells derived from pulp of deciduous teeth

• Published in: Archives of oral biology 2018-12, Vol.96, p.201-207
• Main Authors: Aghajani, Farzaneh, Kazemnejad, Somaieh, Hooshmand, Tabassom, Ghaempanah, Zahra, Zarnani, Amir-Hassan
• Format: Article
• Language: English
• Subjects: Biomarkers - metabolism; Cell Differentiation; Cell Proliferation; Cells, Cultured; Child; Dental pulp; Dental Pulp - cytology; Dentistry; Female; Flow Cytometry; Human deciduous teeth; Humans; Immunophenotyping - methods; Male; Mesenchymal stem cells; Osteogenesis - physiology; Osteogenic differentiation; Proliferation; Real-Time Polymerase Chain Reaction; Stage-Specific Embryonic Antigens - metabolism; Stem Cells - cytology; Tooth, Deciduous
• ISSN: 0003-9969; 1879-1506
• DOI: 10.1016/j.archoralbio.2018.09.014

•No specific marker has been yet found for isolating stem cells of deciduous teeth pulp.•SHEDs had high differentiation potentials even in the unsorted cells.•SSEA-4-positive cells had a slightly better osteogenic potential.•SSEA-4 may not be a specific marker of superior differentiation capacity. Despite the increased interest in stem cells isolated from remnant pulp of deciduous teeth, no specific marker has been yet established for them. The present study aimed to investigate whether SSEA-4 (stage-specific embryonic antigen) would be a suitable marker to isolate stem cells from Human Exfoliated Deciduous teeth (SHEDs) in order to increase its differentiation potential toward osseous tissue. The SHEDs were isolated and the expression patterns of mesenchymal, hematopoietic and embryonic stem cell markers were assessed. The cells were then divided into two groups of SSEA-4(+) and unsorted SHEDs and the cell proliferation rate and population-doubling-time (PDT) were calculated. Subsequently, the differentiation potentials were examined through alizarin-red staining and Quantitative real time-PCR (qRT-PCR). Isolated cells were spindle-shaped with a high expression of mesenchymal stem cell markers and weak expression of hematopoietic markers. The mean expression of Oct-4 was 68.77%±1.28. Despite similar proliferation rates between SSEA-4(+) and unsorted SHEDs, because of differences in the shape of the growth curves, PDT was lower in unsorted SHEDs (P = 0.2 × 10−4). Alizarin-red staining showed similar calcium deposition in both groups. Upon differentiation, the expression of osteocalcin was higher in unsorted SHEDs (P = 0.043), while, the expression of alkaline phosphatase was lower (P