Volume 13, Issue 50 (3-2023)
NCMBJ 2023, 13(50): 103-114 |
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Nazm Bojnordi M, Gholipourmalekabadi M, Ghasemi Hamidabadi H, Keshavarz H. Trans Differentiation of Human Dental Pulp Stem Cells into Dopaminergic neurons via Neurosphere Pulp Stem Cells into Dopaminergic neurons. NCMBJ 2023; 13 (50) :103-114
URL: http://ncmbjpiau.ir/article-1-1600-en.html
URL: http://ncmbjpiau.ir/article-1-1600-en.html
Emergency medicine department, Rasool-e-akram hospital, Iran University of Medical Sciences (IUMS), Tehran, Iran
Abstract: (772 Views)
Aim and Background: Human dental pulp stem cells (hDPSCs) that closely common origin with ectomesenchymal cells. could be applied as the source of stem cells for treatment of some neurodegenerative diseases. The hDPSCc are capable of differentiating into dopaminergic neurons in presence of appropriate neurotrophic factors and neurosphere technique with regard to fact This study was carried out to assess the differentiation potential of hDPSCc into dopaminergic neurons using neurosphere technique.
Materials and Methods: In this study, hDPSCs were isolated from the third molar using mechanical-enzymatic methods. The cells were harvested in a proper condition medium and differentiated into precursor neuronal stem cells and then consequently differentiated into dopaminergic cells in the presence of neuronal induction media including BDNF and GDNF. Cells in each groups were evaluated by immunocytochemistry and RT-PCR assays. In addition, the amount of dopamine released was measured by HPLC.
Results: Cells treated with differentiation inducer factors exhibited remarkable changes in phenotypic characteristics and neuron like cells were observed. Molecular results showed that pluripotent genes such as Nestin , Sox2 expression level gradually reduced in the early stages of differentiation process. However, after treatment with neuronal inducers, genes expression level involved in neurogenesis/dopaminergic like DAT ,PITX3, ,Nurr1 ,TH PITX3 in treated group increased significantly compared to control group. Moreover, immune cytochemistry assay indicated that cells expressing tyrosine hydroxylase (TH) were also detected in treated group. In addition, dopamine release was significantly increased in treated group (P < 0.05).
Conclusion: These research findings confirm that the hDPSCs can be differentiated into dopaminergic neurons via neurosphere technique and appropriate inducers.
Materials and Methods: In this study, hDPSCs were isolated from the third molar using mechanical-enzymatic methods. The cells were harvested in a proper condition medium and differentiated into precursor neuronal stem cells and then consequently differentiated into dopaminergic cells in the presence of neuronal induction media including BDNF and GDNF. Cells in each groups were evaluated by immunocytochemistry and RT-PCR assays. In addition, the amount of dopamine released was measured by HPLC.
Results: Cells treated with differentiation inducer factors exhibited remarkable changes in phenotypic characteristics and neuron like cells were observed. Molecular results showed that pluripotent genes such as Nestin , Sox2 expression level gradually reduced in the early stages of differentiation process. However, after treatment with neuronal inducers, genes expression level involved in neurogenesis/dopaminergic like DAT ,PITX3, ,Nurr1 ,TH PITX3 in treated group increased significantly compared to control group. Moreover, immune cytochemistry assay indicated that cells expressing tyrosine hydroxylase (TH) were also detected in treated group. In addition, dopamine release was significantly increased in treated group (P < 0.05).
Conclusion: These research findings confirm that the hDPSCs can be differentiated into dopaminergic neurons via neurosphere technique and appropriate inducers.
Keywords: Human dental pulp stem cells, neurosphere, neuroprogenitors, dopaminergic neurons, differentiation, Iau Science.
Type of Study: Research Article |
Subject:
Physiology
Received: 2022/11/7 | Accepted: 2022/11/30 | Published: 2023/03/21
Received: 2022/11/7 | Accepted: 2022/11/30 | Published: 2023/03/21
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