Abstract
Neural stem cells (NSCs) have a high potency for differentiation to neurons and glial cells for replacement of damaged cells and paracrine effects for the regeneration and remyelination of host axons. Dental pulp is known to have a potential to differentiate into neural-like cells; therefore, dental pulp may be used as an autologous cell source for neural repair. In this study, we selectively expanded stem cells from human dental pulp in an initial culture using NSC media under xeno- and serum-free conditions. At the initial step of primary culture, human dental pulp was divided into two groups according to the culture media: 10% fetal bovine serum medium group (FBS group) and NSC culture medium group (NSC group). In the NSC group relative to the FBS group, the expression of NSC markers and the concentrations of leukemia inhibitory factor, nerve growth factor, and stem cell factor were higher, although their expression levels were lower than those of human fetal NSCs. The transplanted cells of the NSC group survived well within the normal brain and injured spinal cord of rats and expressed nestin and Sox2. Under the xeno- and serum-free conditions, autologous human dental pulp-derived stem cells might prove useful for clinical cell-based therapies to repair damaged neural tissues.
Highlights
Stem cell-based therapies using neural stem cells (NSCs) are considered to be one of the most promising strategies for treatment of the lesions of the central and peripheral nervous systems [1, 2]
In the experimental group (NSC group), cells were cultured in NSC media: DMEM/F12 supplemented with 1% P/S, 20 ng/mL basic fibroblast growth factor, 20 ng/mL epidermal growth factor (EGF, R&D Systems), N2 (Invitrogen), and B27 (Invitrogen), under the xeno- and serum-free conditions
Two control cell lines were used to compare the cell metabolic activity and quantitative real-time polymerase chain reaction analysis: bone-marrow-derived mesenchymal stem cells (MSCs) (BMMSCs, from ATCC) which were cultured in DMEM/F12 medium supplemented with 10% fetal bovine serum (FBS), 1% P/S, and human fetal brain-derived NSCs which were cultured in DMEM/F12 medium supplemented with 1% P/S, bFGF (20 ng/mL), leukemia inhibitory factor (10 ng/mL, Sigma), and N2
Summary
Stem cell-based therapies using neural stem cells (NSCs) are considered to be one of the most promising strategies for treatment of the lesions of the central and peripheral nervous systems [1, 2]. Developed induced pluripotent stem cells (iPSCs) and direct reprogrammed NSCs [7,8,9] have potential risks of viral integration, tumor formation, and genomic instability which remain hurdles to clinical translation [10]. Stem cells within dental pulp (i.e., dental pulp stem cells: DPSCs) have a high potential for proliferation and differentiation into neural-like cells and as such might be a good source for neural regeneration [13]. Previous studies have demonstrated successful differentiation of human dental pulp-derived stem cells into neural-like cells in both in vitro and in vivo conditions [14,15,16,17], and other studies, have found that neural-like
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