Single-Cell RNA Sequencing Analysis of Human Dental Pulp Stem Cell and Human Periodontal Ligament Stem Cell

Abstract

Introduction: Many studies have investigated the transcriptome profiles of dental stem cells for regenerative medicine. However, such studies use bulk RNA and do not consider cell-level heterogeneity. Here, we investigated the characteristics and heterogeneity of human dental pulp stem cells (hDPSCs) and human periodontal ligament stem cells (hPDLSCs) at the single-cell level and examined the differences between them. Methods: hDPSCs and hPDLSCs were obtained from caries-free premolars (n = 2). Single-cell RNA sequencing of hDPSCs and hPDLSCs was performed using a Single Emulsion Systems (Dolomite Microfluidics, Royston, UK) droplet microfluidic device and the Illumina NextSeq550 system (Illumina, San Diego, CA). Data alignment was performed using STAR v2.4.0, and further analysis was performed using the Seurat, singleR, and clusterProfiler packages in R (R Foundation for Statistical Computing, Vienna, Austria). Results: hDPSCs and hPDLSCs were separated into 3 clusters. hDPSCs mainly exhibited osteogenic and neurogenic cell populations. The main populations of hPDLSCs comprised osteogenic and myofibroblastic populations. hPDLSCs showed high "scores" for osteogenic gene expression, whereas hDPSCs had high neurogenic and endogenic scores. Conclusions: Single-cell RNA sequencing of hDPSCs and hPDLSCs revealed that the genes were expressed in specific clusters. The results of these analyses can be used as reference databases and valuable resources for further research in dental therapeutics.