Induction of periodontal ligament-derived mesenchymal stromal cell-like cells from human induced pluripotent stem cells

Abstract

Introduction

Periodontal disease is a common oral infection which affects the tooth-supportive tissues directly. Considering the limitation of present regenerative treatments for severe periodontal cases, cytotherapies have been gradually introduced. Human periodontal ligament-derived mesenchymal stromal cells (hPDLMSCs), while identified as one of the promising cell sources for periodontal regenerative therapy, still hold some problems in the clinical application especially their limited life span. To solve the problems, human induced pluripotent stem cells (hiPSCs) are taken into consideration as a robust supply for hPDLMSCs.

Methods

The induction of hPDLMSCs was performed based on the generation of neural crest-like cells (NCLCs) from hiPSCs. Fibronectin and laminin were tested as coating materials for NCLCs differentiation when following previous protocol, and the characteristics of induced cells were identified by flow cytometry and RT-qPCR for evaluating the induction efficiency. Subsequently, selected dental ectoderm signaling-related cytokines were applied for hPDLMSCs induction for 14 days, and dental mesenchyme-related genes, dental follicle-related genes and hPDL-related genes were tested by RT-qPCR for the evaluation of differentiation.

Results

Compared to the 58% in laminin-coated condition, fibronectin-coated condition had a higher induction efficiency of CD271 ^high cells as 86% after 8-day induction, while the mesenchymal potential of induced NCLCs was similar between two coating materials.

It was shown that the gene expressions of dental mesenchyme, dental follicles and hPDL cells were significantly enhanced with the stimulation of the combination with fibroblast growth factor 8b (FGF8b), FGF2, and bone morphogenetic protein 4 (BMP4).

Conclusion

FN coating was more effective in NCLCs induction, and the FGF8b+FGF2+BMP4 growth factor cocktail was effective in hPDLMSC-like cell generation. These findings underscored the likely regenerative potential of hiPSCs as an applicable and promising curative strategy for periodontal diseases.

Highlights

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  The matrix-dependent active derivation of neural crest-like cells (NCLCs) from human iPSCs.

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  FGF8b, BMP4, FGF2 treatment induced iNCLCs toward dental mesenchymal stem cells differentiation.

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  Induced MSCs with the stimulation of growth factors expressed hPDLs-related markers including POSTN, S100A4.