Exploring the Effects of Long-Term Cultivation on the Properties of Stem Cells From Apical Papilla (SCAP)

Objectives: The cultivation of Stem cells from apical papilla (SCAP) under suitable conditions has demonstrated remarkable potential for proliferation and differentiation. As a dependable cell source for diverse regenerative therapies, human SCAP has gained attention. Nonetheless, the impact of aging on the properties of stem cells remains largely unknown. Thus, the objective of this study is to investigate the effects of extended cultivation on SCAP and to identify indications of cell senescence.
Methods: Healthy patients' routinely extracted third molars were utilized to obtain SCAP tissue, from which stem cells were isolated and cultured under standard conditions (DMEM supplemented with 10% fetal bovine serum) for an extended period in vitro. The SCAPs were cultivated for nearly four months, spanning from the first to the twentieth passage. To identify any signs of aging, cells from early (1st-3rd), medium (10th-12th), and late (18th-20th) passages were harvested and evaluated for cell proliferation and apoptosis. Additionally, telomerase and beta-galactosidase enzymatic activity, known to be associated with cell senescence, were thoroughly investigated. Finally, an MTT test was administered.
Results: We successfully isolated and expanded SCAPs in vitro. Our findings indicate that there is no statistically significant reduction in the proliferative capacity of stem cells in early and late passages (p>0.05). We observed a slightly increased number of apoptotic cells in late passage stem cells. However, the enzymatic activity of telomerase and beta-galactosidase remained relatively stable in both early and late passages, with no statistically significant difference observed in our stem cell lineages.
Conclusions: Changes that occur during long-term cultivation are typically considered when intending to use stem cells for research or therapy. Nevertheless, our results indicate that there is no statistically significant variation in the proliferative characteristics and enzymatic activity of SCAPs between early and late passages.