Neurogenic differentiation of human dental pulp stem cells by optogenetics stimulation

Niyazi, M. and Zibaii, M.I. and Chavoshinezhad, S. and Hamidabadi, H.G. and Dargahi, L. and Bojnordi, M.N. and Alizadeh, R. and Heravi, M. and Karimi, H. and Hosseini, M. and Sadeghi Malvajerdi, E. and Seyednazari, M. (2020) Neurogenic differentiation of human dental pulp stem cells by optogenetics stimulation. Journal of Chemical Neuroanatomy, 109.

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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 � 104 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. © 2020

Item Type: Article
Additional Information: cited By 0
Uncontrolled Keywords: doublecortin; lentivirus vector; microtubule associated protein 2; nestin; opsin, adult; Article; blue light; cell density; cell differentiation; cell isolation; cell lineage; cell shape; cell stimulation; cell structure; cell viability; controlled study; dental pulp stem cell; enzyme assisted extraction; human; human cell; immunocytochemistry; MTT assay; nerve cell; optogenetics; phase contrast microscopy; priority journal; third molar
Subjects: QU Biochemistry. Cell Biology and Genetics
WU Dentistry. Oral Surgery
Depositing User: eprints admin
Date Deposited: 03 Oct 2020 10:29
Last Modified: 03 Oct 2020 10:29

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