Neuronal differentiation of dental pulp stem cells from human permanent and deciduous teeth following coculture with rat auditory brainstem slices

Abstract

Sensorineural hearing loss is a common disability found worldwide which is associated with a degeneration of spiral ganglion neurons (SGN). It is a challenge to restore SGN due to the permanent degeneration and viability of SGN is requisite for patients to receive an advantage from hearing aid devices. Human dental pulp stem cells (DPSC) and stem cells from human exfoliated deciduous teeth (SHED) are self-renewing stem cells that originate from the neural crest during development. These stem cells have a high potential for neuronal differentiation. This is primarily due to their multilineage differentiation potential and their relative ease of access. Previously, we have shown the ability of these stem cell types to differentiate into spiral ganglion neuron-like cells. In this study, we induced the cells into neural precursor cells (NPC) and cocultured with auditory brainstem slice (ABS) encompassing cochlear nucleus by the Stoppini method. We also investigated their ability to differentiate after 2 weeks and 4 weeks in coculture. Neuronal differentiation of DPSC-NPC and SHED-NPC was higher expression of specific markers to SGN, TrkB, and Gata3, compared to monoculture. The cells also highly expressed synaptic vesicle protein (SV2A) and exhibited intracellular calcium oscillations. Our findings demonstrated the possibility of using DPSCs and SHEDs as an autologous stem cell-based therapy for sensorineural hearing loss patients. K E Y W O R D S auditory, brainstem slice (ABS), deciduous teeth (SHED), human dental pulp stem cell (DPSC), spiral ganglion neuron (SGN), stem cell from human exfoliated

Potential of resveratrol in enrichment of neural progenitor-like cell induction of human stem cells from apical papilla

“…The differentiated cells exhibited a neuronal shape which positively detected neural cytoskeletal proteins such as β-III tubulin and neurofilament [12,13,20] or neuronal cell-specific markers such as Nestin, CD133, glial fibrillary acid protein (GFAP), MAP-2, enolase, and synaptophysin [11,15,20,21]. Second are functional characteristics involving functional neuronal networks, intercellular communication [40,41], and intracellular signaling cascades [12,23]. Calcium ions are internalized into neurons to modulate vesicular neurotransmitter releasing [42].…”

“…In order to identify potential of neuronal differentiation, we evaluated calcium influx which is an indicator for neurotransmitter transmission. The intracellular calcium assessment was described in previous study [23]. The specimens were incubated with 3 μM Fluo-3 AM (Invitrogen) and 0.08% pluronic acid (Invitrogen) in DMEM/F-12, 100 U/mL penicillin, and 100 μM/mL streptomycin at 37°C for 60 min.…”

Potential of resveratrol in enrichment of neural progenitor-like cells induction of human stem cells from apical papilla

“…The differentiated cells exhibited neuronal shaped which positively detected neural cytoskeletal proteins such as β-III tubulin, neuro lament (12,13,20) or neuronal cell speci c markers such as Nestin, CD133, glial brillary acid protein (GFAP), MAP-2, enolase, synaptophysin (11,15,20,21). Secondly, functional characteristics involving functional neuronal networks, intercellular communication (40,41), and intracellular signaling cascades (12,23). Calcium ions are internalized into neurons to modulate vesicular neurotransmitter releasing (42).…”

“…In order to identify potential of neuronal differentiation, we evaluated calcium in ux which is an indicator for neurotransmitter transmission. The intracellular calcium assessment was described in previous study (23). The specimens were incubated with 3 µM Fluo-3 AM (Invitrogen) and 0.08% pluronic acid (Invitrogen) in DMEM/F-12, 100 U/mL Penicillin, 100 μM/mL Streptomycin at 37 o C for 60 minutes.…”

Potential of resveratrol in enrichment of neural progenitor-like cells induction of human stem cells from apical papilla

“…In order to identify potential of neuronal differentiation, we evaluated calcium in ux which is an indicator for neurotransmitter transmission. The intracellular calcium assessment was described in previous study (23). The specimens were incubated with 3 µM Fluo-3 AM (Invitrogen) and 0.08%…”