Transcriptome analysis of basic fibroblast growth factor treated stem cells isolated from human exfoliated deciduous teeth

Nunthawan Nowwarote,Jeeranan Manokawinchoke,Kiattipan Kanjana,Benjamin P.J Fournier,Waleerat Sukarawan,Thanaphum Osathanon

doi:10.1016/j.heliyon.2020.e04246

Abstract

BackgroundBasic fibroblast growth factor (bFGF) regulates cell proliferation, migration, and differentiation in various cell types. The aim of the present study was to determine the bFGF target genes in stem cells isolated from human exfoliated deciduous teeth (SHEDs). MethodsCells were isolated from pulp tissue obtained from exfoliated deciduous teeth. Mesenchymal stem cell surface markers and the differentiation potential toward adipogenic and neurogenic lineages were characterized. The bFGF-treated SHED transcriptome was examined using a high throughput RNA sequencing technique. The mRNA and protein expression of selected genes were evaluated using real-time polymerase chain reaction and immunofluorescence staining, respectively. Cell cycle analysis was performed by flow cytometry. The colony forming unit number was also examined. ResultsThe isolated cells expressed CD44, CD90, CD105, but not CD45. The upregulation of adipogenic and neurogenic marker genes was observed after culturing cells in the appropriate induction medium. Transcriptome analysis of the bFGF treated cells revealed that the upregulated genes were in the cell cycle related pathways, while the downregulated genes were in the extracellular matrix related pathways. Correspondingly, bFGF induced MKI67 mRNA expression and Ki67 protein expression. Furthermore, bFGF treatment significantly decreased the G0/G1, but increased the G2/M, population in SHEDs. Colony formation was markedly increased in the bFGF treated group and was attenuated by pretreating the cells with FGFR or PI3K inhibitors. ConclusionbFGF controls cell cycle progression in SHEDs. Thus, it can be used to amplify cell number to obtain the amount of cells required for regenerative treatments.

Highlights

Stem cells from exfoliated deciduous teeth (SHEDs) are considered to be an alternative source of mesenchymal stem cells because they can be non-invasively isolated from the dental pulp tissue of exfoliated primary teeth
Transplanting SHEDs in murine calvarial defects resulted in a similar amount of bone regeneration compared with human dental pulp stem cells and human bone marrow mesenchymal stem cells [6]
The present study found that Basic fibroblast growth factor (bFGF) promoted SHED colony forming unit number via the FGFR and PI3K pathways

Summary

Introduction

Stem cells from exfoliated deciduous teeth (SHEDs) are considered to be an alternative source of mesenchymal stem cells because they can be non-invasively isolated from the dental pulp tissue of exfoliated primary teeth. After maintaining SHEDs in the appropriate induction medium, the cells differentiated into cell lineages derived from ectoderm, mesoderm, and endoderm [1,2]. In addition to their multi-differentiation potency, SHEDs exhibit immunomodulatory properties. SHEDs combined with collagen scaffolds promoted dental pulp tissue formation and newly deposited tubular dentin in subcutaneously implanted root canals [7]. Based on these findings, SHEDs have been proposed as an alternative mesenchymal stem cell source for regenerative therapy, especially in the dental field. It can be used to amplify cell number to obtain the amount of cells required for regenerative treatments

Objectives

Methods

Results

Discussion

Conclusion