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从人乳恒牙中分离干细胞并将其分化为螺旋神经节神经元样细胞。

Differentiation of stem cells from human deciduous and permanent teeth into spiral ganglion neuron-like cells.

机构信息

Department of Anatomy, Faculty of Science, Mahidol University, Bangkok, Thailand.

Department of Pediatric Dentistry, Faculty of Dentistry, Mahidol University, Bangkok, Thailand.

出版信息

Arch Oral Biol. 2018 Apr;88:34-41. doi: 10.1016/j.archoralbio.2018.01.011. Epub 2018 Feb 2.

Abstract

OBJECTIVE

Stem cells from pulp tissue are a promising cell-based therapy for neurodegenerative patients based on their origin in the neural crest. The aim of this study was to differentiate and evaluate the ability of human dental pulp stem cells from permanent teeth (DPSC) and stem cells from human exfoliated deciduous teeth (SHED) to differentiate into spiral ganglion neurons.

DESIGN

After isolation and characterization of mesenchymal stem cell properties, DPSC and SHED were treated with the neurotrophins brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and glial cell-derived neurotrophic factor (GDNF). The differentiation was identified by immunostaining and qRT-PCR analysis of neuronal markers and measuring intracellular calcium activity.

RESULTS

After 2 weeks of induction, morphological changes were observed in both DPSC and SHED. The differentiated cells expressed neuron-specific class III beta-tubulin, GATA binding protein 3 (GATA3) and tropomyosin receptor kinase B, protein markers of spiral ganglion neurons. These cells also showed upregulation of the genes encoding these proteins, namely GATA3 and neurotrophic receptor tyrosine kinase 2. Intracellular calcium dynamics that reflect neurotransmitter release were observed in differentiated DPSC and SHED.

CONCLUSION

These results demonstrate that dental pulp stem cells from permanent and deciduous teeth can differentiate into spiral ganglion neuron-like cells.

摘要

目的

牙髓组织中的干细胞来源于神经嵴,是一种很有前途的神经退行性疾病的基于细胞的治疗方法。本研究的目的是分化和评估人恒牙牙髓干细胞(DPSC)和人脱落乳牙牙髓干细胞(SHED)分化为螺旋神经节神经元的能力。

设计

在分离和鉴定间充质干细胞特性后,用神经营养因子脑源性神经营养因子(BDNF)、神经营养因子-3(NT-3)和胶质细胞源性神经营养因子(GDNF)处理 DPSC 和 SHED。通过神经元标志物的免疫染色和 qRT-PCR 分析以及测量细胞内钙活性来鉴定分化。

结果

诱导 2 周后,DPSC 和 SHED 均观察到形态学变化。分化细胞表达神经元特异性 III 型β-微管蛋白、GATA 结合蛋白 3(GATA3)和原肌球蛋白受体激酶 B,这是螺旋神经节神经元的蛋白标志物。这些细胞还表现出编码这些蛋白的基因 GATA3 和神经营养受体酪氨酸激酶 2 的上调。在分化的 DPSC 和 SHED 中观察到反映神经递质释放的细胞内钙动力学。

结论

这些结果表明,来自恒牙和乳牙的牙髓干细胞可以分化为螺旋神经节神经元样细胞。

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