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In vitro neural/glial differentiation potential of periodontal ligament stem cells.

Li X, Gong P, Liao D - Arch Med Sci (2010)

Bottom Line: We conclude that PDLSCs have neural/glial differentiation potential in vitro and that neural/glial differentiation can be induced in PDLSCs if suitable protocols are followed.We also found that supplementing the growth medium with suitable growth factors is more effective than applying chemicals alone.While nerve growth factor is more effective than platelet-derived growth factor for inducing neural/glial differentiation in PDLSCs, pre-induction of PDLSCs with dimethyl sulphoxide yields better results than those obtained with all-trans-retinoic acid.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, China.

ABSTRACT

Introduction: It is known that periodontal ligament stem cells (PDLSCs) can differentiate into cementoblast-like cells, adipocytes and collagen-forming cells. However, whether PDLSCs are able to differentiate into Schwann cells and which method is best for their neural induction remain unknown. We attempted to determine whether PDLSCs possessed the potential for neural differentiation in vitro.

Materials and methods: We isolated and multiplied PDLSCs from periodontal ligaments obtained from the teeth (n = 24) of 8-month-old beagle dogs. Four protocols with different chemicals and growth factors were adopted to induce the PDLSCs to differentiate into Schwann cells. Immunochemistry, RT-PCR and qRT-PCR were performed to investigate the in vitro neural differentiation potential of PDLSCs.

Results: We compared the 4 different protocols and showed that all 4 protocols could successfully induce PDLSCs to express nestin, GFAP and S100, markers for Schwann cells. Further, qRT-PCR revealed relative differences in the expression levels of these 3 genes in differentiated PDLSCs obtained by different protocols.

Conclusions: We conclude that PDLSCs have neural/glial differentiation potential in vitro and that neural/glial differentiation can be induced in PDLSCs if suitable protocols are followed. We also found that supplementing the growth medium with suitable growth factors is more effective than applying chemicals alone. While nerve growth factor is more effective than platelet-derived growth factor for inducing neural/glial differentiation in PDLSCs, pre-induction of PDLSCs with dimethyl sulphoxide yields better results than those obtained with all-trans-retinoic acid.

No MeSH data available.


Related in: MedlinePlus

In the graph, bar height represents the gene expression relative quantity (mean ± SEM). For GFAP and S100 gene expression relative quantity, statistic analyses show no statistical differences between group A and B, C and D, respectively (P > 0.05), but group C and D have significantly statistical differences as compered with group A and B (P < 0.05); for Nestin gene expression relative quantity, there are no statistical differences between group A and B (P > 0.05), but there are statistical differences between group C and D (P < 0.05), and the results of group C or D are significantly higher than those of group A or B (P < 0.05)
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Figure 4: In the graph, bar height represents the gene expression relative quantity (mean ± SEM). For GFAP and S100 gene expression relative quantity, statistic analyses show no statistical differences between group A and B, C and D, respectively (P > 0.05), but group C and D have significantly statistical differences as compered with group A and B (P < 0.05); for Nestin gene expression relative quantity, there are no statistical differences between group A and B (P > 0.05), but there are statistical differences between group C and D (P < 0.05), and the results of group C or D are significantly higher than those of group A or B (P < 0.05)

Mentions: To confirm the immunocytochemistry results, RT-PCR was performed, and the result is shown in Figure 3. All dPDLSCs from all 4 protocols were positive for S100, nestin and GFAP, but the uPDLSCs were negative for S100, nestin and GFAP. Subsequently, we performed real-time PCR to consolidate the findings. For S100 and GFAP, statistical analyses showed no statistically significant differences between protocols A and B, or C and D, respectively (P > 0.05). However, the results of protocols C and D showed statistically significant differences as compared with those of protocols A and B (P < 0.05). Gene expression of S100 in protocols C and D was higher than that in protocols A and B (Tables II and III). For nestin gene expression, there were no statistically significant differences between protocols A and B (P > 0.05), but there were differences between protocols C and D (P < 0.05). All the results of protocols C and D were significantly higher than those of protocols A and B (P < 0.05), and nestin gene expression was the highest in protocol C, closely followed by protocol D (Table IV, Figure 4).


In vitro neural/glial differentiation potential of periodontal ligament stem cells.

Li X, Gong P, Liao D - Arch Med Sci (2010)

In the graph, bar height represents the gene expression relative quantity (mean ± SEM). For GFAP and S100 gene expression relative quantity, statistic analyses show no statistical differences between group A and B, C and D, respectively (P > 0.05), but group C and D have significantly statistical differences as compered with group A and B (P < 0.05); for Nestin gene expression relative quantity, there are no statistical differences between group A and B (P > 0.05), but there are statistical differences between group C and D (P < 0.05), and the results of group C or D are significantly higher than those of group A or B (P < 0.05)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
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getmorefigures.php?uid=PMC3298334&req=5

Figure 4: In the graph, bar height represents the gene expression relative quantity (mean ± SEM). For GFAP and S100 gene expression relative quantity, statistic analyses show no statistical differences between group A and B, C and D, respectively (P > 0.05), but group C and D have significantly statistical differences as compered with group A and B (P < 0.05); for Nestin gene expression relative quantity, there are no statistical differences between group A and B (P > 0.05), but there are statistical differences between group C and D (P < 0.05), and the results of group C or D are significantly higher than those of group A or B (P < 0.05)
Mentions: To confirm the immunocytochemistry results, RT-PCR was performed, and the result is shown in Figure 3. All dPDLSCs from all 4 protocols were positive for S100, nestin and GFAP, but the uPDLSCs were negative for S100, nestin and GFAP. Subsequently, we performed real-time PCR to consolidate the findings. For S100 and GFAP, statistical analyses showed no statistically significant differences between protocols A and B, or C and D, respectively (P > 0.05). However, the results of protocols C and D showed statistically significant differences as compared with those of protocols A and B (P < 0.05). Gene expression of S100 in protocols C and D was higher than that in protocols A and B (Tables II and III). For nestin gene expression, there were no statistically significant differences between protocols A and B (P > 0.05), but there were differences between protocols C and D (P < 0.05). All the results of protocols C and D were significantly higher than those of protocols A and B (P < 0.05), and nestin gene expression was the highest in protocol C, closely followed by protocol D (Table IV, Figure 4).

Bottom Line: We conclude that PDLSCs have neural/glial differentiation potential in vitro and that neural/glial differentiation can be induced in PDLSCs if suitable protocols are followed.We also found that supplementing the growth medium with suitable growth factors is more effective than applying chemicals alone.While nerve growth factor is more effective than platelet-derived growth factor for inducing neural/glial differentiation in PDLSCs, pre-induction of PDLSCs with dimethyl sulphoxide yields better results than those obtained with all-trans-retinoic acid.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, China.

ABSTRACT

Introduction: It is known that periodontal ligament stem cells (PDLSCs) can differentiate into cementoblast-like cells, adipocytes and collagen-forming cells. However, whether PDLSCs are able to differentiate into Schwann cells and which method is best for their neural induction remain unknown. We attempted to determine whether PDLSCs possessed the potential for neural differentiation in vitro.

Materials and methods: We isolated and multiplied PDLSCs from periodontal ligaments obtained from the teeth (n = 24) of 8-month-old beagle dogs. Four protocols with different chemicals and growth factors were adopted to induce the PDLSCs to differentiate into Schwann cells. Immunochemistry, RT-PCR and qRT-PCR were performed to investigate the in vitro neural differentiation potential of PDLSCs.

Results: We compared the 4 different protocols and showed that all 4 protocols could successfully induce PDLSCs to express nestin, GFAP and S100, markers for Schwann cells. Further, qRT-PCR revealed relative differences in the expression levels of these 3 genes in differentiated PDLSCs obtained by different protocols.

Conclusions: We conclude that PDLSCs have neural/glial differentiation potential in vitro and that neural/glial differentiation can be induced in PDLSCs if suitable protocols are followed. We also found that supplementing the growth medium with suitable growth factors is more effective than applying chemicals alone. While nerve growth factor is more effective than platelet-derived growth factor for inducing neural/glial differentiation in PDLSCs, pre-induction of PDLSCs with dimethyl sulphoxide yields better results than those obtained with all-trans-retinoic acid.

No MeSH data available.


Related in: MedlinePlus