Establishing and characterizing human stem cells from the apical papilla immortalized by hTERT gene transfer

Stem cells from the apical papilla (SCAPs) are promising candidates for regenerative endodontic treatment and tissue regeneration in general. However, harvesting enough cells from the limited apical papilla tissue is difficult, and the cells lose their primary phenotype over many passages. To get over these challenges, we immortalized human SCAPs with lentiviruses overexpressing human telomerase reverse transcriptase (hTERT). Human immortalized SCAPs (hiSCAPs) exhibited long-term proliferative activity without tumorigenic potential. Cells also expressed mesenchymal and progenitor biomarkers and exhibited multiple differentiation potentials. Interestingly, hiSCAPs gained a stronger potential for osteogenic differentiation than the primary cells. To further investigate whether hiSCAPs could become prospective seed cells in bone tissue engineering, in vitro and in vivo studies were performed, and the results indicated that hiSCAPs exhibited strong osteogenic differentiation ability after infection with recombinant adenoviruses expressing BMP9 (AdBMP9). In addition, we revealed that BMP9 could upregulate ALK1 and BMPRII, leading to an increase in phosphorylated Smad1 to induce the osteogenic differentiation of hiSCAPs. These results support the application of hiSCAPs in tissue engineering/regeneration schemes as a stable stem cell source for osteogenic differentiation and biomineralization, which could be further used in stem cell-based clinical therapies.

Automated summary

Stem cells from the apical papilla (SCAPs) are a promising candidate for tissue regeneration and regenerative endodontic treatment. However, obtaining enough cells from the limited apical papilla tissue and maintaining their primary phenotype is challenging. This study successfully immortalized human SCAPs, allowing for long-term proliferation and retaining mesenchymal and progenitor biomarkers. The immortalized cells showed greater potential for osteogenic differentiation compared to primary cells, making them a potential source for bone tissue engineering.