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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 markers in DPCs. (a) Cells were cultured in the presence or absence of CIP (10 μg/mL) for 72 h. The cells at the early passages were used as an untreated control at 0 h. Expression of CD133 and procollagen type I were analyzed by immunofluorescence staining. Scale bar is 50 μm. (b) Time-dependent effects of CIP treatment on the expression of stem cell markers were determined. Cells were cultured in the presence or absence of CIP (10 μg/mL) for 0–72 h. The levels of procollagen type I, CD133, integrin β1, and ALDH1A1 were determined by western blot analysis. Blots were reprobed with β-actin to confirm equal loading. The immunoblot signals were quantified by densitometry and the mean data from independent experiments were normalized to the results. The data represent the means of four independent samples ± SD. ∗P < 0.05 versus untreated control at 0 h. (c) Cells were treated with CIP (0–10 μg/mL) for 72 h. After indicated treatment, levels of procollagen type I, CD133, integrin β1, and ALDH1A1 were analyzed by western blot. β-actin was served as the loading control. The immunoblot signals were quantified by densitometry and the 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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fig3: Effects of CIP on stem cell markers in DPCs. (a) Cells were cultured in the presence or absence of CIP (10 μg/mL) for 72 h. The cells at the early passages were used as an untreated control at 0 h. Expression of CD133 and procollagen type I were analyzed by immunofluorescence staining. Scale bar is 50 μm. (b) Time-dependent effects of CIP treatment on the expression of stem cell markers were determined. Cells were cultured in the presence or absence of CIP (10 μg/mL) for 0–72 h. The levels of procollagen type I, CD133, integrin β1, and ALDH1A1 were determined by western blot analysis. Blots were reprobed with β-actin to confirm equal loading. The immunoblot signals were quantified by densitometry and the mean data from independent experiments were normalized to the results. The data represent the means of four independent samples ± SD. ∗P < 0.05 versus untreated control at 0 h. (c) Cells were treated with CIP (0–10 μg/mL) for 72 h. After indicated treatment, levels of procollagen type I, CD133, integrin β1, and ALDH1A1 were analyzed by western blot. β-actin was served as the loading control. The immunoblot signals were quantified by densitometry and the 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: Having shown that culture of the DPCs caused the spontaneous decline of stem cell-like phenotypes, we next clarify the mentioned conception by detecting stem cell markers in such cells. Because the CD133 and procollagen type I expressions have been recognized as the dermal papilla cell and fibroblast indicators, respectively [11, 23, 24], we analyzed the expression of such proteins in the DPCs treated with CIP at the concentrations of 10 μg/mL for 72 h and the control cells. Immunocytochemistry showed that the expression of CD133 with CIP was suppressed in the DPCs cells after being cultivated for 72 h in comparison to that of control cells (DPCs at the early passages at 0 h, Figure 3(a)). Treatment of the cells with CIP could dramatically prevent such a loss of CD133 expression in the cells (Figure 3(a)). These results suggested that CIP preserve the stemness of DPCs during culture. Also, the expression level of fibroblast marker procollagen type I was significantly increased in the untreated DPCs cells at 72 h, whereas the increase in fibroblast marker could be prevented by CIP at the concentration of 10 μg/mL (Figure 3(a)).


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 markers in DPCs. (a) Cells were cultured in the presence or absence of CIP (10 μg/mL) for 72 h. The cells at the early passages were used as an untreated control at 0 h. Expression of CD133 and procollagen type I were analyzed by immunofluorescence staining. Scale bar is 50 μm. (b) Time-dependent effects of CIP treatment on the expression of stem cell markers were determined. Cells were cultured in the presence or absence of CIP (10 μg/mL) for 0–72 h. The levels of procollagen type I, CD133, integrin β1, and ALDH1A1 were determined by western blot analysis. Blots were reprobed with β-actin to confirm equal loading. The immunoblot signals were quantified by densitometry and the mean data from independent experiments were normalized to the results. The data represent the means of four independent samples ± SD. ∗P < 0.05 versus untreated control at 0 h. (c) Cells were treated with CIP (0–10 μg/mL) for 72 h. After indicated treatment, levels of procollagen type I, CD133, integrin β1, and ALDH1A1 were analyzed by western blot. β-actin was served as the loading control. The immunoblot signals were quantified by densitometry and the 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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Related In: Results  -  Collection

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fig3: Effects of CIP on stem cell markers in DPCs. (a) Cells were cultured in the presence or absence of CIP (10 μg/mL) for 72 h. The cells at the early passages were used as an untreated control at 0 h. Expression of CD133 and procollagen type I were analyzed by immunofluorescence staining. Scale bar is 50 μm. (b) Time-dependent effects of CIP treatment on the expression of stem cell markers were determined. Cells were cultured in the presence or absence of CIP (10 μg/mL) for 0–72 h. The levels of procollagen type I, CD133, integrin β1, and ALDH1A1 were determined by western blot analysis. Blots were reprobed with β-actin to confirm equal loading. The immunoblot signals were quantified by densitometry and the mean data from independent experiments were normalized to the results. The data represent the means of four independent samples ± SD. ∗P < 0.05 versus untreated control at 0 h. (c) Cells were treated with CIP (0–10 μg/mL) for 72 h. After indicated treatment, levels of procollagen type I, CD133, integrin β1, and ALDH1A1 were analyzed by western blot. β-actin was served as the loading control. The immunoblot signals were quantified by densitometry and the 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: Having shown that culture of the DPCs caused the spontaneous decline of stem cell-like phenotypes, we next clarify the mentioned conception by detecting stem cell markers in such cells. Because the CD133 and procollagen type I expressions have been recognized as the dermal papilla cell and fibroblast indicators, respectively [11, 23, 24], we analyzed the expression of such proteins in the DPCs treated with CIP at the concentrations of 10 μg/mL for 72 h and the control cells. Immunocytochemistry showed that the expression of CD133 with CIP was suppressed in the DPCs cells after being cultivated for 72 h in comparison to that of control cells (DPCs at the early passages at 0 h, Figure 3(a)). Treatment of the cells with CIP could dramatically prevent such a loss of CD133 expression in the cells (Figure 3(a)). These results suggested that CIP preserve the stemness of DPCs during culture. Also, the expression level of fibroblast marker procollagen type I was significantly increased in the untreated DPCs cells at 72 h, whereas the increase in fibroblast marker could be prevented by CIP at the concentration of 10 μg/mL (Figure 3(a)).

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