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Neural crest stem cells from dental tissues: a new hope for dental and neural regeneration.

Ibarretxe G, Crende O, Aurrekoetxea M, García-Murga V, Etxaniz J, Unda F - Stem Cells Int (2012)

Bottom Line: However, endogenous adult sources of neural stem cells present major drawbacks, such as their scarcity and complicated obtention.In this context, EMSCs from dental tissues emerge as good alternative candidates, since they are preserved in adult human individuals, and retain both high proliferation ability and a neural-like phenotype in vitro.We will finally review some of the latest research featuring experimental approaches and benefits of dental stem cell therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology and Histology, Faculty of Medicine and Dentistry, University of the Basque Country (UPV/EHU), 48940 Bizkaia, Leioa, Spain.

ABSTRACT
Several stem cell sources persist in the adult human body, which opens the doors to both allogeneic and autologous cell therapies. Tooth tissues have proven to be a surprisingly rich and accessible source of neural crest-derived ectomesenchymal stem cells (EMSCs), which may be employed to repair disease-affected oral tissues in advanced regenerative dentistry. Additionally, one area of medicine that demands intensive research on new sources of stem cells is nervous system regeneration, since this constitutes a therapeutic hope for patients affected by highly invalidating conditions such as spinal cord injury, stroke, or neurodegenerative diseases. However, endogenous adult sources of neural stem cells present major drawbacks, such as their scarcity and complicated obtention. In this context, EMSCs from dental tissues emerge as good alternative candidates, since they are preserved in adult human individuals, and retain both high proliferation ability and a neural-like phenotype in vitro. In this paper, we discuss some important aspects of tissue regeneration by cell therapy and point out some advantages that EMSCs provide for dental and neural regeneration. We will finally review some of the latest research featuring experimental approaches and benefits of dental stem cell therapy.

No MeSH data available.


Related in: MedlinePlus

Dental EMSCs express neural differentiation and pluripotency markers and can acquire a prominent neural-like morphology in vitro. Dental EMSCs isolated from dental pulp (DPSCs) form clonogenic adherent colonies (a), which present Nestin+ immunoreactivity (b) and from which equally Nestin+ migrating cells spread to eventually bring the culture plate to full confluency (c). Dental EMSCs also express pluripotency markers such as Oct-4, in the absence of any genetic or pharmacological manipulation (d). The cellular morphology and proliferation rates of dental EMSCs vary depending on the presence of FBS in the culture medium. DPSCs proliferate slowly in the absence of serum. Cells cultured without serum are equally Nestin+ but display very variable morphologies, including the appearance of cells with striking neuron-like shape, that show very thin and long cytoplasmic processes, resembling dendrites and axons (f, g). When DPSC are expanded for 3 weeks with 10% FBS, following another 3 weeks of serum deprivation, a sheet of nerve-like tissue is formed (h). Times after seeding: (a) 1 week; (e) 3 weeks; (h) 6 weeks (3 + 3); (b–d; f–g) double merged images of Nestin (green) and Oct-4 (red) immunolabeled cells, with DAPI (blue) introduced as a nuclear counterstain. Scale bars: 50 μm.
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fig2: Dental EMSCs express neural differentiation and pluripotency markers and can acquire a prominent neural-like morphology in vitro. Dental EMSCs isolated from dental pulp (DPSCs) form clonogenic adherent colonies (a), which present Nestin+ immunoreactivity (b) and from which equally Nestin+ migrating cells spread to eventually bring the culture plate to full confluency (c). Dental EMSCs also express pluripotency markers such as Oct-4, in the absence of any genetic or pharmacological manipulation (d). The cellular morphology and proliferation rates of dental EMSCs vary depending on the presence of FBS in the culture medium. DPSCs proliferate slowly in the absence of serum. Cells cultured without serum are equally Nestin+ but display very variable morphologies, including the appearance of cells with striking neuron-like shape, that show very thin and long cytoplasmic processes, resembling dendrites and axons (f, g). When DPSC are expanded for 3 weeks with 10% FBS, following another 3 weeks of serum deprivation, a sheet of nerve-like tissue is formed (h). Times after seeding: (a) 1 week; (e) 3 weeks; (h) 6 weeks (3 + 3); (b–d; f–g) double merged images of Nestin (green) and Oct-4 (red) immunolabeled cells, with DAPI (blue) introduced as a nuclear counterstain. Scale bars: 50 μm.

Mentions: The amount of cells that can be obtained from a healthy human molar tooth pulp ranges between 500.000 and 2 million, which may seem quite modest. However, it is estimated that between 0.2% and 0.7% of the cells plated after pulp dissociation represent true colony-forming dental EMSCs, also referred to as dental pulp stem cells (DPSC) [39]. In our experience with these dental pulp cultures, when placed in a culture medium specific for MSC, nonstem cells deadhere and only adherent dental EMSCs remain. These EMSCs rapidly generate Oct-4+/Vimentin+/Nestin+ clonogenic colonies. After 5 days in culture, each of the colonies may show around 40–50 cells on average, and some peripheral cells with fibroblastic migratory shape, showing big lamellipodia, begin to spread apart of the colony cell mass (Figures 2(a)-2(b)). Then, a significant change is observed, notably depending on the absence or presence of fetal bovine serum (FBS) in the culture medium. Cells placed in 10% FBS continue to proliferate at high rate and can be maintained in this condition for very long periods, over 4–6 months, while preserving Oct-4+/Vimentin+/Nestin+ immunoreactivity (Figures 2(c)-2(d)). We estimate that, in the presence of FBS, about 1000 plated EMSCs are capable of bringing a 6-well culture plate area to full confluence (roughly 1 million cells) in the course of merely 2 weeks. Thus, it seems reasonable to say that, although the number of EMSCs that can be obtained from a single tooth piece is indeed small, their high proliferative capacity in vitro makes dental tissues very promising alternatives to provide sufficient amounts of EMSCs, even for clinical purposes.


Neural crest stem cells from dental tissues: a new hope for dental and neural regeneration.

Ibarretxe G, Crende O, Aurrekoetxea M, García-Murga V, Etxaniz J, Unda F - Stem Cells Int (2012)

Dental EMSCs express neural differentiation and pluripotency markers and can acquire a prominent neural-like morphology in vitro. Dental EMSCs isolated from dental pulp (DPSCs) form clonogenic adherent colonies (a), which present Nestin+ immunoreactivity (b) and from which equally Nestin+ migrating cells spread to eventually bring the culture plate to full confluency (c). Dental EMSCs also express pluripotency markers such as Oct-4, in the absence of any genetic or pharmacological manipulation (d). The cellular morphology and proliferation rates of dental EMSCs vary depending on the presence of FBS in the culture medium. DPSCs proliferate slowly in the absence of serum. Cells cultured without serum are equally Nestin+ but display very variable morphologies, including the appearance of cells with striking neuron-like shape, that show very thin and long cytoplasmic processes, resembling dendrites and axons (f, g). When DPSC are expanded for 3 weeks with 10% FBS, following another 3 weeks of serum deprivation, a sheet of nerve-like tissue is formed (h). Times after seeding: (a) 1 week; (e) 3 weeks; (h) 6 weeks (3 + 3); (b–d; f–g) double merged images of Nestin (green) and Oct-4 (red) immunolabeled cells, with DAPI (blue) introduced as a nuclear counterstain. Scale bars: 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
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fig2: Dental EMSCs express neural differentiation and pluripotency markers and can acquire a prominent neural-like morphology in vitro. Dental EMSCs isolated from dental pulp (DPSCs) form clonogenic adherent colonies (a), which present Nestin+ immunoreactivity (b) and from which equally Nestin+ migrating cells spread to eventually bring the culture plate to full confluency (c). Dental EMSCs also express pluripotency markers such as Oct-4, in the absence of any genetic or pharmacological manipulation (d). The cellular morphology and proliferation rates of dental EMSCs vary depending on the presence of FBS in the culture medium. DPSCs proliferate slowly in the absence of serum. Cells cultured without serum are equally Nestin+ but display very variable morphologies, including the appearance of cells with striking neuron-like shape, that show very thin and long cytoplasmic processes, resembling dendrites and axons (f, g). When DPSC are expanded for 3 weeks with 10% FBS, following another 3 weeks of serum deprivation, a sheet of nerve-like tissue is formed (h). Times after seeding: (a) 1 week; (e) 3 weeks; (h) 6 weeks (3 + 3); (b–d; f–g) double merged images of Nestin (green) and Oct-4 (red) immunolabeled cells, with DAPI (blue) introduced as a nuclear counterstain. Scale bars: 50 μm.
Mentions: The amount of cells that can be obtained from a healthy human molar tooth pulp ranges between 500.000 and 2 million, which may seem quite modest. However, it is estimated that between 0.2% and 0.7% of the cells plated after pulp dissociation represent true colony-forming dental EMSCs, also referred to as dental pulp stem cells (DPSC) [39]. In our experience with these dental pulp cultures, when placed in a culture medium specific for MSC, nonstem cells deadhere and only adherent dental EMSCs remain. These EMSCs rapidly generate Oct-4+/Vimentin+/Nestin+ clonogenic colonies. After 5 days in culture, each of the colonies may show around 40–50 cells on average, and some peripheral cells with fibroblastic migratory shape, showing big lamellipodia, begin to spread apart of the colony cell mass (Figures 2(a)-2(b)). Then, a significant change is observed, notably depending on the absence or presence of fetal bovine serum (FBS) in the culture medium. Cells placed in 10% FBS continue to proliferate at high rate and can be maintained in this condition for very long periods, over 4–6 months, while preserving Oct-4+/Vimentin+/Nestin+ immunoreactivity (Figures 2(c)-2(d)). We estimate that, in the presence of FBS, about 1000 plated EMSCs are capable of bringing a 6-well culture plate area to full confluence (roughly 1 million cells) in the course of merely 2 weeks. Thus, it seems reasonable to say that, although the number of EMSCs that can be obtained from a single tooth piece is indeed small, their high proliferative capacity in vitro makes dental tissues very promising alternatives to provide sufficient amounts of EMSCs, even for clinical purposes.

Bottom Line: However, endogenous adult sources of neural stem cells present major drawbacks, such as their scarcity and complicated obtention.In this context, EMSCs from dental tissues emerge as good alternative candidates, since they are preserved in adult human individuals, and retain both high proliferation ability and a neural-like phenotype in vitro.We will finally review some of the latest research featuring experimental approaches and benefits of dental stem cell therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology and Histology, Faculty of Medicine and Dentistry, University of the Basque Country (UPV/EHU), 48940 Bizkaia, Leioa, Spain.

ABSTRACT
Several stem cell sources persist in the adult human body, which opens the doors to both allogeneic and autologous cell therapies. Tooth tissues have proven to be a surprisingly rich and accessible source of neural crest-derived ectomesenchymal stem cells (EMSCs), which may be employed to repair disease-affected oral tissues in advanced regenerative dentistry. Additionally, one area of medicine that demands intensive research on new sources of stem cells is nervous system regeneration, since this constitutes a therapeutic hope for patients affected by highly invalidating conditions such as spinal cord injury, stroke, or neurodegenerative diseases. However, endogenous adult sources of neural stem cells present major drawbacks, such as their scarcity and complicated obtention. In this context, EMSCs from dental tissues emerge as good alternative candidates, since they are preserved in adult human individuals, and retain both high proliferation ability and a neural-like phenotype in vitro. In this paper, we discuss some important aspects of tissue regeneration by cell therapy and point out some advantages that EMSCs provide for dental and neural regeneration. We will finally review some of the latest research featuring experimental approaches and benefits of dental stem cell therapy.

No MeSH data available.


Related in: MedlinePlus