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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 characteristics in DPCs. (a) Cells were treated with CIP (0–10 μg/mL) for various times (0–72 h). After indicated treatment, morphology of DPCs was observed. Scale bar is 100 μm. (b) Aggregation behavior of cells was determined after indicated treatment for 72 h. Scale bar is 100 μm. ((c)-(d)) Aggregation size and aggregation number were determined by image analyzer. The data represent the means of four independent samples ± SD. ∗P < 0.05 versus untreated control. ((e)-(f)) Cells were cultured in the presence or absence of CIP (10 μg/mL) for 72 h and serum-starved for 24 h. After serum-starvation, cells were incubated with complete media for 12 h. The cells at the early passages (passages 2-3) without serum-starvation were also used as an untreated control at 0 h. Cell cycle distribution was determined by PI staining and flow cytometry. The data represent the means of four independent samples ± SD. ∗P < 0.05 versus untreated control at 0 h; #P < 0.05 versus untreated control at 72 h.
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fig2: Effects of CIP on stem cell-like characteristics in DPCs. (a) Cells were treated with CIP (0–10 μg/mL) for various times (0–72 h). After indicated treatment, morphology of DPCs was observed. Scale bar is 100 μm. (b) Aggregation behavior of cells was determined after indicated treatment for 72 h. Scale bar is 100 μm. ((c)-(d)) Aggregation size and aggregation number were determined by image analyzer. The data represent the means of four independent samples ± SD. ∗P < 0.05 versus untreated control. ((e)-(f)) Cells were cultured in the presence or absence of CIP (10 μg/mL) for 72 h and serum-starved for 24 h. After serum-starvation, cells were incubated with complete media for 12 h. The cells at the early passages (passages 2-3) without serum-starvation were also used as an untreated control at 0 h. Cell cycle distribution was determined by PI staining and flow cytometry. The data represent the means of four independent samples ± SD. ∗P < 0.05 versus untreated control at 0 h; #P < 0.05 versus untreated control at 72 h.

Mentions: DPCs have been reported to function as multipotent stem cells and the stemness of DPCs was linked to their ability to induce hair follicles [10–12]. However, DPCs lose their hair follicle inductive ability during culture [4, 6–8]. We found that, after culturing the DPCs for 5 days, the shape and appearance of DPCs are spontaneously altered toward fibroblast-like morphology. The primitive DPCs usually appearing as spindle-shaped cells changed to flat multipolar cells with elongated shapes (Figure 2(a)). Besides, the DPCs at the beginning showed an aggregative growth pattern in culture and such pattern was lost during the extended time of culturing. In order to test whether CIP affects the change in morphology of these DPCs, the cells were treated with CIP at the concentrations of 0–10 μg/mL for 0–72 h, and morphology of the cells as well as aggregative pattern was determined. Figure 2(a) shows that most of untreated control cells exhibited fibroblast-like morphology at 48 and 72 h. Meanwhile, the morphology of CIP treated cells remained unaltered (Figure 2(a)). Because the hair follicle inductive property of the DPCs has been shown to relate with their aggregate behaviors [22], we further investigated the effect of CIP treatments on the aggregative growth pattern of the cells. The DPCs at early passages (passages 2-3) were cultured in the presence or absence of CIP for 72 h and the aggregate size and number were determined. Figures 2(b), 2(c), and 2(d) show that CIP at the concentration of 5 and 10 μg/mL significantly increased the size as well as the number of cell aggregation in comparison to those of untreated control at 72 h.


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 characteristics in DPCs. (a) Cells were treated with CIP (0–10 μg/mL) for various times (0–72 h). After indicated treatment, morphology of DPCs was observed. Scale bar is 100 μm. (b) Aggregation behavior of cells was determined after indicated treatment for 72 h. Scale bar is 100 μm. ((c)-(d)) Aggregation size and aggregation number were determined by image analyzer. The data represent the means of four independent samples ± SD. ∗P < 0.05 versus untreated control. ((e)-(f)) Cells were cultured in the presence or absence of CIP (10 μg/mL) for 72 h and serum-starved for 24 h. After serum-starvation, cells were incubated with complete media for 12 h. The cells at the early passages (passages 2-3) without serum-starvation were also used as an untreated control at 0 h. Cell cycle distribution was determined by PI staining and flow cytometry. The data represent the means of four independent samples ± SD. ∗P < 0.05 versus untreated control at 0 h; #P < 0.05 versus untreated control at 72 h.
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Related In: Results  -  Collection

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fig2: Effects of CIP on stem cell-like characteristics in DPCs. (a) Cells were treated with CIP (0–10 μg/mL) for various times (0–72 h). After indicated treatment, morphology of DPCs was observed. Scale bar is 100 μm. (b) Aggregation behavior of cells was determined after indicated treatment for 72 h. Scale bar is 100 μm. ((c)-(d)) Aggregation size and aggregation number were determined by image analyzer. The data represent the means of four independent samples ± SD. ∗P < 0.05 versus untreated control. ((e)-(f)) Cells were cultured in the presence or absence of CIP (10 μg/mL) for 72 h and serum-starved for 24 h. After serum-starvation, cells were incubated with complete media for 12 h. The cells at the early passages (passages 2-3) without serum-starvation were also used as an untreated control at 0 h. Cell cycle distribution was determined by PI staining and flow cytometry. The data represent the means of four independent samples ± SD. ∗P < 0.05 versus untreated control at 0 h; #P < 0.05 versus untreated control at 72 h.
Mentions: DPCs have been reported to function as multipotent stem cells and the stemness of DPCs was linked to their ability to induce hair follicles [10–12]. However, DPCs lose their hair follicle inductive ability during culture [4, 6–8]. We found that, after culturing the DPCs for 5 days, the shape and appearance of DPCs are spontaneously altered toward fibroblast-like morphology. The primitive DPCs usually appearing as spindle-shaped cells changed to flat multipolar cells with elongated shapes (Figure 2(a)). Besides, the DPCs at the beginning showed an aggregative growth pattern in culture and such pattern was lost during the extended time of culturing. In order to test whether CIP affects the change in morphology of these DPCs, the cells were treated with CIP at the concentrations of 0–10 μg/mL for 0–72 h, and morphology of the cells as well as aggregative pattern was determined. Figure 2(a) shows that most of untreated control cells exhibited fibroblast-like morphology at 48 and 72 h. Meanwhile, the morphology of CIP treated cells remained unaltered (Figure 2(a)). Because the hair follicle inductive property of the DPCs has been shown to relate with their aggregate behaviors [22], we further investigated the effect of CIP treatments on the aggregative growth pattern of the cells. The DPCs at early passages (passages 2-3) were cultured in the presence or absence of CIP for 72 h and the aggregate size and number were determined. Figures 2(b), 2(c), and 2(d) show that CIP at the concentration of 5 and 10 μg/mL significantly increased the size as well as the number of cell aggregation in comparison to those of untreated control at 72 h.

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