Effect of ciliary neurotrophic factor on neural differentiation of stem cells of human exfoliated deciduous teeth

Sujuan Zeng,Wanghong Zhao,Yan Shu,Wenyan Huang,Janak L Pathak,Hongbing Guan,Yunyang Li,Lingling Zhang,Lihong Ge,Xuedan Zhao

doi:10.1186/s13036-020-00251-4

Abstract

The stem cells of human exfoliated deciduous teeth (SHEDs) are considered to be one of the main sources of seed cells in stem cell therapy. The aim of this study was to examine the effect of ciliary neurotrophic factor (CNTF) on neurogenic differentiation of SHEDs. With the consent of parents, SHEDs from 6 to 8 year old children were isolated and cultured. The mesenchymal stemness and the potential of multidirectional (adipogenic and osteogenic) differentiation for the isolated SHEDs were firstly determined. The effect of CNTF on specific neurogenic differentiation of SHEDs was then examined by detecting the expression of marker genes and proteins via RT-PCR, immunoblotting, and immunofluorescence microscopy. The isolated SHEDs expressed specific surface markers of mesenchymal stem cells, and their potential of osteogenic and adipogenic differentiation were confirmed. CNTF promoted the differentiation of SHEDs into neuron-like cells with a high expression of acetylcholine transferase (CHAT), a marker of cholinergic neurons. The expression of other neuron markers including nestin, microtubule-associated protein 2 (MAP 2), and β-tublin III was also detected. Interestingly, the expression of neurogenic markers was maintained at a high level after neurogenic induction. SHEDs can be induced by CNTF to differentiate into cholinergic neuron-like cells under appropriate culture conditions. Our findings have laid a foundation for future use of SHEDs to treat neurological diseases.

Highlights

Stem cells are those with the potential to differentiate into different types of cells in the body
By using Alizarin red to stain calcium, we found that stem cells of human exfoliated deciduous teeth (SHEDs) cultured with osteogenic medium for 14 days had apparent formation of mineralized nodules as compared to those cultured in control medium which resulted in little staining (Fig. 3A)
We found that ciliary neurotrophic factor (CNTF) treatment could facilitate neurogenic differentiation of SHEDs

Summary

Introduction

Stem cells are those with the potential to differentiate into different types of cells in the body. Many types of human stem cells have been derived from oral cavity, including dental pulp stem cells (DPSCs), stem cells of human exfoliated deciduous teeth (SHEDs), apical papillary stem cells, and periodontal ligament stem cells [3]. These stem cells are all mesenchymal origin. The neuronal differentiation capability of SHEDs has been reported to be stronger than those of DPSCs and bone marrow stem cells (BMSCs) [6]. Protocols have been developed to promote the differentiation of stem cells including SHEDs into neuron-like cells [12, 13]. Given that SHEDs are available from the clinic with minimal ethical concerns, the potential of SHEDs in stem cell therapy for neurological and other diseases is of particular interest

Objectives

Methods

Results

Conclusion