Neurogenic differentiation of dental pulp stem cells to neuron-like cells in dopaminergic and motor neuronal inductive media

被引:73
作者
Chang, Chia-Chieh [1 ,2 ]
Chang, Kai-Chun [1 ]
Tsai, Shang-Jye [1 ,3 ]
Chang, Hao-Hueng [1 ,4 ,5 ]
Lin, Chun-Pin [1 ,4 ,5 ]
机构
[1] Natl Taiwan Univ, Sch Dent, Grad Inst Clin Dent, Taipei 10048, Taiwan
[2] Taitung Anim Propagat Stn COA LRI, Taitung 954, Taiwan
[3] Cardinal Tien Hosp, Yung Ho Branch, Dept Dent, New Taipei City, Taiwan
[4] Natl Taiwan Univ, Sch Dent, Dept Dent, Taipei 10764, Taiwan
[5] Natl Taiwan Univ Hosp, Dept Dent, Taipei, Taiwan
关键词
dental pulp stem cells; dopaminergic neuronal inductive media; motor neuronal inductive media; neuron-like cells; BONE-MARROW; IN-VITRO; MICROARRAY ANALYSIS; NERVOUS-SYSTEM; RETINOIC ACID; STROMAL CELLS; TISSUE-REPAIR; CULTURE; TRANSPLANTATION; EXPRESSION;
D O I
10.1016/j.jfma.2014.09.003
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Background/Purpose: Dental pulp stem cells (DPSCs) have been proposed as a promising source of stem cells in nerve regeneration due to their close embryonic origin and ease of harvest. The aim of this study was to evaluate the efficacy of dopaminergic and motor neuronal inductive media on transdifferentiation of human DPSCs (hDPSCs) into neuron-like cells. Methods: Isolation, cultivation, and identification of hDPSCs were performed with morphological analyses and flow cytometry. The proliferation potential of DPSCs was evaluated with an XTT [(2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide)] assay. Media for the induction of dopaminergic and spinal motor neuronal differentiation were prepared. The efficacy of neural induction was evaluated by detecting the expression of neuron cell-specific cell markers in DPSCs by immunocytochemistry and quantitative real-time reverse transcription polymerase chain reaction (RT-PCR). Results: In the XTT assay, there was a 2.6-or 2-fold decrease in DPSCs cultured in dopaminergic or motor neuronal inductive media, respectively. The proportions of beta III-tubulin (beta III-tub), glial fibrillary acidic protein (GFAP), and oligodendrocyte (O1)-positive cells were significantly higher in DPSCs cultured in both neuronal inductive media compared with those cultured in control media. Furthermore, hDPSC-derived dopaminergic and spinal motor neuron cells after induction expressed a higher density of neuron cell markers than those before induction. Conclusion: These findings suggest that in response to the neuronal inductive stimuli, a greater proportion of DPSCs stop proliferation and acquire a phenotype resembling mature neurons. Such neural crest- derived adult DPSCs may provide an alternative stem cell source for therapy- based treatments of neuronal disorders and injury. Copyright (C) 2014, Elsevier Taiwan LLC & Formosan Medical Association. All rights reserved.
引用
收藏
页码:956 / 965
页数:10
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