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Ciprofloxacin Improves the Stemness of Human Dermal Papilla Cells.

Kiratipaiboon C, Tengamnuay P, Chanvorachote P - Stem Cells Int (2015)

Bottom Line: We found that ciprofloxacin exerted its effect through ATP-dependent tyrosine kinase/glycogen synthase kinase3β dependent mechanism which in turn upregulated β-catenin.The effects of ciprofloxacin in preserving stem cell features were confirmed in the primary dermal papilla cells directly obtained from human hair follicles.Together, these results revealed a novel application of ciprofloxacin for stem cell maintenance and provided the underlying mechanisms that are responsible for the stemness in dermal papilla cells.

View Article: PubMed Central - PubMed

Affiliation: Pharmaceutical Technology (International) Program, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand.

ABSTRACT
Improvement in the expansion method of adult stem cells may augment their use in regenerative therapy. Using human dermal papilla cell line as well as primary dermal papilla cells as model systems, the present study demonstrated that ciprofloxacin treatment could prevent the loss of stemness during culture. Clonogenicity and stem cell markers of dermal papilla cells were shown to gradually decrease in the culture in a time-dependent manner. Treatment of the cells with nontoxic concentrations of ciprofloxacin could maintain both stem cell morphology and clonogenicity, as well as all stem cells markers. We found that ciprofloxacin exerted its effect through ATP-dependent tyrosine kinase/glycogen synthase kinase3β dependent mechanism which in turn upregulated β-catenin. Besides, ciprofloxacin was shown to induce epithelial-mesenchymal transition in DPCs as the transcription factors ZEB1 and Snail were significantly increased. Furthermore, the self-renewal proteins of Wnt/β-catenin pathway, namely, Nanog and Oct-4 were significantly upregulated in the ciprofloxacin-treated cells. The effects of ciprofloxacin in preserving stem cell features were confirmed in the primary dermal papilla cells directly obtained from human hair follicles. Together, these results revealed a novel application of ciprofloxacin for stem cell maintenance and provided the underlying mechanisms that are responsible for the stemness in dermal papilla cells.

No MeSH data available.


Related in: MedlinePlus

Effects of CIP on stem cell-like phenotypes in primary human DPCs. (a) Cells were treated with CIP (0–10 μg/mL) for 24 h. Cytotoxicity was determined by MTT assay. The data represent the means of four independent triplicate samples ± SD. (b) After indicated treatment, mode of cell death was examined by Hoechst 33342/PI costaining assay. Scale bar is 100 μm. ((c)–(e)) Cells were treated with CIP (0–10 μg/mL) for 72 h. After indicated treatment, the levels of stem cell markers (procollagen type I, CD133, integrin β1, and ALDH1A1), Wnt/β-catenin signaling (Akt, p-Akt, GSK3β, p-GSK3β, and β-catenin), and EMT transcription factors (ZEB1, Slug, and Snail) were determined by western blot analysis, respectively. β-actin was served as the loading control. The immunoblot signals were quantified by densitometry and mean data from independent experiments were normalized to the results. The data represent the means of four independent samples ± SD. ∗P < 0.05 and ∗∗P < 0.01 versus untreated control.
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fig5: Effects of CIP on stem cell-like phenotypes in primary human DPCs. (a) Cells were treated with CIP (0–10 μg/mL) for 24 h. Cytotoxicity was determined by MTT assay. The data represent the means of four independent triplicate samples ± SD. (b) After indicated treatment, mode of cell death was examined by Hoechst 33342/PI costaining assay. Scale bar is 100 μm. ((c)–(e)) Cells were treated with CIP (0–10 μg/mL) for 72 h. After indicated treatment, the levels of stem cell markers (procollagen type I, CD133, integrin β1, and ALDH1A1), Wnt/β-catenin signaling (Akt, p-Akt, GSK3β, p-GSK3β, and β-catenin), and EMT transcription factors (ZEB1, Slug, and Snail) were determined by western blot analysis, respectively. β-actin was served as the loading control. The immunoblot signals were quantified by densitometry and mean data from independent experiments were normalized to the results. The data represent the means of four independent samples ± SD. ∗P < 0.05 and ∗∗P < 0.01 versus untreated control.

Mentions: To determine the extent to which primary human DPCs will respond to the CIP treatment by the same manner, we treated the isolated human DPCs with 0–10 μg/mL CIP and evaluated stem cell characteristics, accordingly. Figures 5(a) and 5(b) indicate that treatment of the cells with 0–10 μg/mL CIP caused no direct cytotoxicity in these cells. We next tested the signature characteristics of stem cells to assess whether CIP sustained the stemness in these primary cells. The cells were cultured in the presence or absence of CIP for 72 h and the expression of stem cells as well as fibroblast markers was determined as described previously. Western blot analysis revealed that the levels of CD133, integrin β1, and ALDH1A1 were significantly increased in response to CIP treatment in a dose-dependent manner, whereas the expression of procollagen type I was significantly suppressed (Figure 5(c)). Furthermore, the Wnt/β-catenin and EMT were evaluated by western blotting. Figure 5(d) shows that treatment of the cells with CIP significantly increased the level of activated Akt level, inactivated GSK3β, and β-catenin. Besides, the EMT transcription factors ZEB1 and Snail were found to be upregulated in response to the treatment (Figure 5(e)). Taken together, these data supported our earlier findings that CIP maintains the stemness of DPCs via β-catenin and EMT.


Ciprofloxacin Improves the Stemness of Human Dermal Papilla Cells.

Kiratipaiboon C, Tengamnuay P, Chanvorachote P - Stem Cells Int (2015)

Effects of CIP on stem cell-like phenotypes in primary human DPCs. (a) Cells were treated with CIP (0–10 μg/mL) for 24 h. Cytotoxicity was determined by MTT assay. The data represent the means of four independent triplicate samples ± SD. (b) After indicated treatment, mode of cell death was examined by Hoechst 33342/PI costaining assay. Scale bar is 100 μm. ((c)–(e)) Cells were treated with CIP (0–10 μg/mL) for 72 h. After indicated treatment, the levels of stem cell markers (procollagen type I, CD133, integrin β1, and ALDH1A1), Wnt/β-catenin signaling (Akt, p-Akt, GSK3β, p-GSK3β, and β-catenin), and EMT transcription factors (ZEB1, Slug, and Snail) were determined by western blot analysis, respectively. β-actin was served as the loading control. The immunoblot signals were quantified by densitometry and mean data from independent experiments were normalized to the results. The data represent the means of four independent samples ± SD. ∗P < 0.05 and ∗∗P < 0.01 versus untreated control.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig5: Effects of CIP on stem cell-like phenotypes in primary human DPCs. (a) Cells were treated with CIP (0–10 μg/mL) for 24 h. Cytotoxicity was determined by MTT assay. The data represent the means of four independent triplicate samples ± SD. (b) After indicated treatment, mode of cell death was examined by Hoechst 33342/PI costaining assay. Scale bar is 100 μm. ((c)–(e)) Cells were treated with CIP (0–10 μg/mL) for 72 h. After indicated treatment, the levels of stem cell markers (procollagen type I, CD133, integrin β1, and ALDH1A1), Wnt/β-catenin signaling (Akt, p-Akt, GSK3β, p-GSK3β, and β-catenin), and EMT transcription factors (ZEB1, Slug, and Snail) were determined by western blot analysis, respectively. β-actin was served as the loading control. The immunoblot signals were quantified by densitometry and mean data from independent experiments were normalized to the results. The data represent the means of four independent samples ± SD. ∗P < 0.05 and ∗∗P < 0.01 versus untreated control.
Mentions: To determine the extent to which primary human DPCs will respond to the CIP treatment by the same manner, we treated the isolated human DPCs with 0–10 μg/mL CIP and evaluated stem cell characteristics, accordingly. Figures 5(a) and 5(b) indicate that treatment of the cells with 0–10 μg/mL CIP caused no direct cytotoxicity in these cells. We next tested the signature characteristics of stem cells to assess whether CIP sustained the stemness in these primary cells. The cells were cultured in the presence or absence of CIP for 72 h and the expression of stem cells as well as fibroblast markers was determined as described previously. Western blot analysis revealed that the levels of CD133, integrin β1, and ALDH1A1 were significantly increased in response to CIP treatment in a dose-dependent manner, whereas the expression of procollagen type I was significantly suppressed (Figure 5(c)). Furthermore, the Wnt/β-catenin and EMT were evaluated by western blotting. Figure 5(d) shows that treatment of the cells with CIP significantly increased the level of activated Akt level, inactivated GSK3β, and β-catenin. Besides, the EMT transcription factors ZEB1 and Snail were found to be upregulated in response to the treatment (Figure 5(e)). Taken together, these data supported our earlier findings that CIP maintains the stemness of DPCs via β-catenin and EMT.

Bottom Line: We found that ciprofloxacin exerted its effect through ATP-dependent tyrosine kinase/glycogen synthase kinase3β dependent mechanism which in turn upregulated β-catenin.The effects of ciprofloxacin in preserving stem cell features were confirmed in the primary dermal papilla cells directly obtained from human hair follicles.Together, these results revealed a novel application of ciprofloxacin for stem cell maintenance and provided the underlying mechanisms that are responsible for the stemness in dermal papilla cells.

View Article: PubMed Central - PubMed

Affiliation: Pharmaceutical Technology (International) Program, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand.

ABSTRACT
Improvement in the expansion method of adult stem cells may augment their use in regenerative therapy. Using human dermal papilla cell line as well as primary dermal papilla cells as model systems, the present study demonstrated that ciprofloxacin treatment could prevent the loss of stemness during culture. Clonogenicity and stem cell markers of dermal papilla cells were shown to gradually decrease in the culture in a time-dependent manner. Treatment of the cells with nontoxic concentrations of ciprofloxacin could maintain both stem cell morphology and clonogenicity, as well as all stem cells markers. We found that ciprofloxacin exerted its effect through ATP-dependent tyrosine kinase/glycogen synthase kinase3β dependent mechanism which in turn upregulated β-catenin. Besides, ciprofloxacin was shown to induce epithelial-mesenchymal transition in DPCs as the transcription factors ZEB1 and Snail were significantly increased. Furthermore, the self-renewal proteins of Wnt/β-catenin pathway, namely, Nanog and Oct-4 were significantly upregulated in the ciprofloxacin-treated cells. The effects of ciprofloxacin in preserving stem cell features were confirmed in the primary dermal papilla cells directly obtained from human hair follicles. Together, these results revealed a novel application of ciprofloxacin for stem cell maintenance and provided the underlying mechanisms that are responsible for the stemness in dermal papilla cells.

No MeSH data available.


Related in: MedlinePlus