Conditioned-medium of stem cells from human exfoliated deciduous teeth prevent apoptosis of neural progenitors.

Purpose: This study aimed to evaluate the neuroprotective ability of the conditioned medium of stem cells from human exfoliated deciduous teeth (CM-SHED) to prevent glutamate-induced apoptosis of neural progenitors. Materials and methods: Neural progenitors were isolated from two-day-old rat brains, and the conditioned medium was obtained from a mesenchymal stem cell SHED. Four groups were examined: neural progenitor cells cultured in neurobasal medium with (N + ) and without (N-) glutamate and glycine, and neural progenitor cells cultured in CM-SHED with (K + ) and without (K-) glutamate and glycine. Results: The expression of GABA A1 receptor (GABAAR1) messenger RNA (mRNA) in neural progenitor measured by real-time quantitative PCR. GABA contents were measured by enzyme-linked immunosorbent assay, whereas the apoptosis markers caspase-3 and 7-aminoactinomycin D were analysed with a Muse® cell analyzer. The viability of neural progenitor cells in the K + group (78.05 %) was higher than the control group N- (73.22 %) and lower in the N + group (68.90 %) than in the control group. The K + group showed the highest GABA content, which significantly differed from that in the other groups, whereas the lowest content was observed in the N + group. The expression level of GABAAR1 mRNA in the K + group was the highest compared to that in the other groups. CM-SHED potently protected the neural progenitors from apoptosis. Conclusions: CM-SHED may effectively prevent glutamate-induced apoptosis of neural progenitors.

Establishing methods for isolation of stem cells from human exfoliated deciduous from carious deciduous teeth.

AIM: This study aims to establish the isolation method of stem cells from pulp tissue of carious deciduous teeth. METHODS: The teeth were soaked in 1% povidone-iodine solution for about 1 min followed by washing in PBS with 1% antibiotic-antimycotic thrice. Dental pulp tissue was removed by extirpation, and then cultivated in the culture medium. Characterization of mesenchymal stem cell (MSC) was carried out using human MSC analysis kit with positive markers CD90, CD73, and CD105, but negative for expressions of CD45, CD34, CD11b, CD19, and HLA-DR. Differentiation capacity of stem cells from human exfoliated deciduous (SHED) was determined by staining with Alizarin S, Alcian Blue, and Oil Red O. RESULTS: There is no contamination after 3 days of culture. SHED derived from dental pulp were expressions of 99.2% of positive marker and 0.3% of the negative marker. At passage 5, SHED was differentiated into osteocyte, chondrocyte, and adipocyte types of cells in the induction medium. CONCLUSION: SHED derived from carious deciduous teeth can be used as a source of stem cell for regenerative medicine.