A transcriptomic analysis of dental pulp stem cell senescence in vitro

Area of Science:

• * Regenerative Medicine
• * Stem Cell Biology
• * Tissue Engineering

Background:

• * Human dental pulp stem cells (hDPSCs) are crucial for regenerative medicine.
• * Autologous hDPSCs are investigated for tissue repair applications.
• * Understanding long-term culture effects is vital for clinical translation.

Purpose of the Study:

• * To evaluate the impact of extended in vitro culture on hDPSCs.
• * To assess changes in multidifferentiation capacity over passages.
• * To identify molecular mechanisms underlying culture-induced senescence.

Main Methods:

• * hDPSCs isolated and cultured for up to 12 generations.
• * Flow cytometry for surface marker analysis (CD34, CD45, CD73, CD90, CD105).
• * Proliferation (CCK-8), senescence (β-Galactosidase), osteogenic (Alizarin Red S), and lipogenic (Oil Red O) assays.
• * RNA sequencing (RNA-seq) to identify differentially expressed genes.

Main Results:

• * Increased passage numbers correlated with higher senescence and reduced proliferation.
• * Osteogenic and lipogenic differentiation potential decreased with extended culture.
• * Stable CD34/CD45 expression contrasted with decreased CD73/CD90/CD105.
• * RNA-seq identified CFH, WNT16, HSD17B2, IDI1, and COL5A3 as potentially involved in senescence.

Conclusions:

• * Prolonged in vitro expansion of hDPSCs induces senescence.
• * Senescence diminishes proliferative and differentiation capabilities.
• * Specific gene expression changes (e.g., CFH, WNT16) may underlie hDPSCs senescence.