IADR Abstract Archives
Differentiation Of Human Dental Pulp Stem Cells Into Neural Lineages
Objectives: The present study characterised and compared a population of adult stem cells derived from human dental pulp (hDPSCs).
Objectives:
1. Investigation of self-renewal capacity of the cell population
2. Induction of neural differentiation
3. MSC characterisation of cells via immunocytochemistry using CD105 and CD73 antibodies and respectively via qPCR using CD105, CD90, SOX2 and COL15A1 at three different time intervals: Day 0 (undifferentiated cells), Day 8, and Day 24 in neural differentiation media
4. Analysis at protein and mRNA levels of neural differentiated cells at three time intervals using TUJ1, S100, SOX10, GFAP, NCAM, MAPT
Methods: Dental pulp was extracted from wisdom teeth of ten patients and subsequently cultured as explants in growth media and in neurobasal media to assess for neuronal and glial markers. Analysis was carried out at mRNA and protein level using qPCR and immunocytochemistry. Furthermore undifferentiated hDPSCs were analysed at mRNA and protein level to evaluate the expression of MSCs and neural markers.
Results: We successfully isolated a population of MSCs from the dental pulp, as confirmed by positive CD105 and CD73 staining and qPCR analysis. Undifferentiated DPSCs spontaneously expressed glial marker S100 independent of passage number. Neural differentiation was confirmed by positive immunocytochemical analysis of TUJ1, GFAP and S100. Quantitvate qPCR analysis showed great expression of NCAM and moderately to low expressions of MAPT and S100 when compared to controls.
Conclusions: The results obtained from the current study indicate that dental pulp is a promising source of MCSs. Induced hDPSCs showed a great expression of NCAM, which plays a role in myelination and remyelination as well in analgesic effect of glial cell–derived neurotrophic factor in neuropathic pain. Spontaneous expression of glial marker S100 was evident in the undifferentiated dental pulp stem cells, thus protein expression as the only evidence of MSC differentiation towards a neuronal phenotype should be prohibited.
Objectives:
1. Investigation of self-renewal capacity of the cell population
2. Induction of neural differentiation
3. MSC characterisation of cells via immunocytochemistry using CD105 and CD73 antibodies and respectively via qPCR using CD105, CD90, SOX2 and COL15A1 at three different time intervals: Day 0 (undifferentiated cells), Day 8, and Day 24 in neural differentiation media
4. Analysis at protein and mRNA levels of neural differentiated cells at three time intervals using TUJ1, S100, SOX10, GFAP, NCAM, MAPT
Methods: Dental pulp was extracted from wisdom teeth of ten patients and subsequently cultured as explants in growth media and in neurobasal media to assess for neuronal and glial markers. Analysis was carried out at mRNA and protein level using qPCR and immunocytochemistry. Furthermore undifferentiated hDPSCs were analysed at mRNA and protein level to evaluate the expression of MSCs and neural markers.
Results: We successfully isolated a population of MSCs from the dental pulp, as confirmed by positive CD105 and CD73 staining and qPCR analysis. Undifferentiated DPSCs spontaneously expressed glial marker S100 independent of passage number. Neural differentiation was confirmed by positive immunocytochemical analysis of TUJ1, GFAP and S100. Quantitvate qPCR analysis showed great expression of NCAM and moderately to low expressions of MAPT and S100 when compared to controls.
Conclusions: The results obtained from the current study indicate that dental pulp is a promising source of MCSs. Induced hDPSCs showed a great expression of NCAM, which plays a role in myelination and remyelination as well in analgesic effect of glial cell–derived neurotrophic factor in neuropathic pain. Spontaneous expression of glial marker S100 was evident in the undifferentiated dental pulp stem cells, thus protein expression as the only evidence of MSC differentiation towards a neuronal phenotype should be prohibited.
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