Abstract
The dental follicle is an ectomesenchymal tissue surrounding the developing tooth germ. Human dental follicle cells (hDFCs) have the capacity to commit to differentiation into multiple cell types. Here we investigated the capacity of hDFCs to differentiate into neural cells and the efficiency of a two-step strategy involving floating neurosphere-like bodies for neural differentiation. Undifferentiated hDFCs showed a spindle-like morphology and were positive for neural markers such as nestin, β-III-tubulin, and S100β. The cellular morphology of several cells was neuronal-like including branched dendrite-like processes and neurites. Next, hDFCs were used for neurosphere formation in serum-free medium containing basic fibroblast growth factor, epidermal growth factor, and B27 supplement. The number of cells with neuronal-like morphology and that were strongly positive for neural markers increased with sphere formation. Gene expression of neural markers also increased in hDFCs with sphere formation. Next, gene expression of neural markers was examined in hDFCs during neuronal differentiation after sphere formation. Expression of Musashi-1 and Musashi-2, MAP2, GFAP, MBP, and SOX10 was upregulated in hDFCs undergoing neuronal differentiation via neurospheres, whereas expression of nestin and β-III-tubulin was downregulated. In conclusion, hDFCs may be another optimal source of neural/glial cells for cell-based therapies to treat neurological diseases.
Highlights
Neurodegenerative disorders are characterized by the loss or atrophy of neurons, leading to functional impairment
Dental follicle tissues were washed in phosphate-buffered saline (PBS), minced with sterilized scalpels, and digested in a solution of 0.1 U/ml collagenase type I and 1 U/ml dispase (Roche, Basel, Switzerland) for 1 h at 37∘C. Human dental follicle cells (hDFCs) attached to 100-mm culture plates and were grown in mesenchymal stem cells (MSCs) growth medium (GM; consisting of MSC basal medium supplemented with fetal bovine serum, l-glutamine, and penicillin/streptomycin; Lonza, Basel, Switzerland) in a humidified sonic, Tokyo, incubator (CO2 Japan) in 5% CO2 incubator MCO-175M; Panain air at 37∘C. hDFCs from the
We examined the potential of hDFCs to differentiate into neuronal-like cells when transferred to appropriate conditions for neuronal differentiation
Summary
Neurodegenerative disorders are characterized by the loss or atrophy of neurons, leading to functional impairment. Various approaches have been proposed to have a beneficial effect on peripheral nerve regeneration, including application of an electric field, administration of neurotrophic factors, and transplantation of stem cells [1,2,3,4]. Implantation of embryonic stem cells, neural stem cells and mesenchymal stem cells (MSCs) is beneficial for peripheral nerve regeneration. MSCs are multipotent stem cells that are capable of differentiating into multiple cell types [5, 6]. The in vitro growth of undifferentiated MSCs, followed by induction of neural cell differentiation and subsequent transplantation, is an important modality for cell therapy to treat neurodegenerative disease [7, 8]. Human bone marrow is generally used as the major source of MSCs to treat neurodegenerative disease [9, 10], MSCs can be derived from all postnatal tissues
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