In Vitro Single-Cell Transcriptomic Profiling of Cultured Stem Cells From Apical Papilla and Dental Pulp Stem Cells:...

Area of Science:

• Stem Cell Biology
• Genomics
• Regenerative Medicine

Background:

• Dental-derived mesenchymal stem cells exhibit variable differentiation potential.
• Nonspecific markers and isolation limitations contribute to this variability.

Purpose of the Study:

• Compare cellular composition of cultured stem cells from apical papilla (SCAPs) and dental pulp (DPSCs) using single-cell RNA sequencing.
• Identify subpopulations and novel markers for SCAP and DPSC heterogeneity.
• Investigate drivers of divergent regenerative behavior.

Main Methods:

• Obtained SCAP and DPSC tissues from human donors.
• Performed single-cell RNA sequencing on cultured cells.
• Utilized Seurat package for dimensionality reduction, clustering, and differential gene expression analysis.
• Applied pseudotime trajectory analysis to infer lineage progression.

Main Results:

• Identified 12 distinct cell clusters shared between SCAP and DPSC cultures.
• Discovered a progenitor-like subset (cluster 9) coexpressing pericyte (NOTCH3, PDGFRB) and mesenchymal markers (MCAM, THY1, DCN).
• Found SCAP-enriched clusters (7 and 9) with higher levels of IGF-associated genes (IGFBP3, IGFBP5).
• Observed DPSC-enriched clusters (10, 11) with stress and apoptotic gene expression.

Conclusions:

• Single-cell transcriptomics reveals significant heterogeneity within cultured SCAPs and DPSCs.
• SCAPs may contain a broader range of proliferative progenitors, indicated by specific gene enrichments.
• Identified NOTCH3 and PDGFRB as potential markers for isolating progenitor-like subsets.
• Findings provide a basis for future research into subpopulation-specific regenerative roles and translational applications.