Neurogenic differentiation of human dental pulp stem cells by optogenetics stimulation.

Autor: Niyazi M; Department of Anatomy & Cell Biology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran. Electronic address: m.niyazi@mazums.ac.ir., Zibaii MI; Laser and Plasma Research Institute-Shahid Beheshti University, Tehran, Iran., Chavoshinezhad S; Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran., Hamidabadi HG; Department of Anatomy & Cell Biology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran; Immunogenetic Research Center, Department of Anatomy & Cell Biology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran. Electronic address: h.ghasemi@mazums.ac.ir., Dargahi L; Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran., Bojnordi MN; Department of Anatomy & Cell Biology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran; Immunogenetic Research Center, Department of Anatomy & Cell Biology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran., Alizadeh R; ENT and Head & Neck Research Center and Department, The Five Senses Institute, Hazrat Rasoul Akram Hospital, Iran University of Medical Sciences, Tehran, Iran., Heravi M; Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran., Karimi H; Laser and Plasma Research Institute-Shahid Beheshti University, Tehran, Iran., Hosseini M; Laser and Plasma Research Institute-Shahid Beheshti University, Tehran, Iran., Sadeghi Malvajerdi E; Laser and Plasma Research Institute-Shahid Beheshti University, Tehran, Iran., Seyednazari M; Laser and Plasma Research Institute-Shahid Beheshti University, Tehran, Iran.
Jazyk: angličtina
Zdroj: Journal of chemical neuroanatomy [J Chem Neuroanat] 2020 Nov; Vol. 109, pp. 101821. Date of Electronic Publication: 2020 Jun 05.
DOI: 10.1016/j.jchemneu.2020.101821
Abstrakt: Introduction: Human dental pulp stem cells (hDPSCs), a promising source for autologous transplantation in regenerative medicine, have been shown to be able to differentiate into neural precursors. Optogenetics is considered as an advanced biological technique in neuroscience which is able to control the activity of genetically modified stem cells by light. The purpose of this study is to investigate the neurogenic differentiation of hDPSCs following optogenetic stimulation.
Methods: The hDPSCs were isolated by mechanical enzymatic digestion from an impacted third molar and cultured in DMEM/F12. The cells were infected with lentiviruses carrying CaMKIIa-hChR2 (H134R). Opsin-expressing hDPSCs were plated at the density of 5 × 10 4 cells/well in 6-well plates and optical stimulation was conducted with blue light (470 nm) pulsing at 15 Hz, 90 % Duty Cycle and 10 mW power for 10 s every 90 minutes, 6 times a day for 5 days. Two control groups including non-opsin-expressing hDPSCs and opsin-expressing hDPSCs with no optical stimulation were also included in the study. A day after last light stimulation, the viability of cells was analyzed by the MTT assay and the morphological changes were examined by phase contrast microscopy. The expression of Nestin, Microtubule-Associated protein 2 (MAP2) and Doublecortin (DCX) were examined by immunocytochemistry.
Results: Human DPSCs expressed the reporter gene, mCherry, 72 hours after lentiviral infection. The result of MTT assay revealed a significant more viability in optical stimulated opsin-expressing hDPSCs as compared with two control groups. Moreover, optical stimulation increased the expression of Nestin, Doublecortin and MAP2 along with morphological changes from spindle shape to neuron-like shape.
Conclusion: Optogenetics stimulation through depolarizing the hDPSCs can increase the cells viability and/or proliferation and also promote the differentiation toward neuron-like cells.
(Copyright © 2020. Published by Elsevier B.V.)
Databáze: MEDLINE
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