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Article Open Access

Hypoxia enhances the wound-healing potential of adipose-derived stem cells in a novel human primary keratinocyte-based scratch assay

  • Authors:
    • Simone Riis
    • Rhonda Newman
    • Hilal Ipek
    • Jens I. Andersen
    • David Kuninger
    • Shayne Boucher
    • Mohan C. Vemuri
    • Cristian P. Pennisi
    • Vladimir Zachar
    • Trine Fink
  • View Affiliations / Copyright

    Affiliations: Laboratory for Stem Cell Research, Department of Health Science and Technology, Aalborg University, Aalborg, Denmark, Thermo Fisher Scientific, Frederick, MD, USA
    Copyright: © Riis et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Pages: 587-594
    |
    Published online on: February 10, 2017
       https://doi.org/10.3892/ijmm.2017.2886
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Abstract

Preclinical studies have suggested that paracrine factors from adipose-derived stem cells (ASCs) promote the healing of chronic wounds, and that the exposure of ASCs to hypoxia enhances their wound healing effect. To aid the translation of these findings into clinical use, robust wound models are necessary to explore each aspect of wound healing. The aspect of re-epithelization is often studied in a scratch assay based on transformed keratinocytes. However, there are concerns regarding the validity of this model, since these cell lines differ from normal keratinocytes, both in terms of proliferative capacity and differentiation, and sensitivity to environmental cues. In this study, the main challenge of using primary keratinocytes to examine the effects of ASCs was identified to be their different requirements for calcium in the culture media. We confirmed that a high calcium content led to morphological and cytoskeletal changes in primary keratinocytes, and demonstrated that a low calcium content compromised the growth of ASCs. We found that it is possible to perform the wound healing assay with primary keratinocytes, if the conditioned media from the ASCs is dialyzed to reduce the calcium concentration. Additionally, using this model of re-epithelization, conditioned media from normoxic ASCs was shown to markedly increase the rate of wound closure by primary keratinocytes, and this effect was significantly enhanced with media from the hypoxia-exposed ASCs. These findings, which are in line with the observations from previous in vivo studies, highlight the validity of this modified assay to investigate the wound healing properties of ASCs in vitro.
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Copy and paste a formatted citation
Spandidos Publications style
Riis S, Newman R, Ipek H, Andersen JI, Kuninger D, Boucher S, Vemuri MC, Pennisi CP, Zachar V, Fink T, Fink T, et al: Hypoxia enhances the wound-healing potential of adipose-derived stem cells in a novel human primary keratinocyte-based scratch assay. Int J Mol Med 39: 587-594, 2017.
APA
Riis, S., Newman, R., Ipek, H., Andersen, J.I., Kuninger, D., Boucher, S. ... Fink, T. (2017). Hypoxia enhances the wound-healing potential of adipose-derived stem cells in a novel human primary keratinocyte-based scratch assay. International Journal of Molecular Medicine, 39, 587-594. https://doi.org/10.3892/ijmm.2017.2886
MLA
Riis, S., Newman, R., Ipek, H., Andersen, J. I., Kuninger, D., Boucher, S., Vemuri, M. C., Pennisi, C. P., Zachar, V., Fink, T."Hypoxia enhances the wound-healing potential of adipose-derived stem cells in a novel human primary keratinocyte-based scratch assay". International Journal of Molecular Medicine 39.3 (2017): 587-594.
Chicago
Riis, S., Newman, R., Ipek, H., Andersen, J. I., Kuninger, D., Boucher, S., Vemuri, M. C., Pennisi, C. P., Zachar, V., Fink, T."Hypoxia enhances the wound-healing potential of adipose-derived stem cells in a novel human primary keratinocyte-based scratch assay". International Journal of Molecular Medicine 39, no. 3 (2017): 587-594. https://doi.org/10.3892/ijmm.2017.2886
Copy and paste a formatted citation
x
Spandidos Publications style
Riis S, Newman R, Ipek H, Andersen JI, Kuninger D, Boucher S, Vemuri MC, Pennisi CP, Zachar V, Fink T, Fink T, et al: Hypoxia enhances the wound-healing potential of adipose-derived stem cells in a novel human primary keratinocyte-based scratch assay. Int J Mol Med 39: 587-594, 2017.
APA
Riis, S., Newman, R., Ipek, H., Andersen, J.I., Kuninger, D., Boucher, S. ... Fink, T. (2017). Hypoxia enhances the wound-healing potential of adipose-derived stem cells in a novel human primary keratinocyte-based scratch assay. International Journal of Molecular Medicine, 39, 587-594. https://doi.org/10.3892/ijmm.2017.2886
MLA
Riis, S., Newman, R., Ipek, H., Andersen, J. I., Kuninger, D., Boucher, S., Vemuri, M. C., Pennisi, C. P., Zachar, V., Fink, T."Hypoxia enhances the wound-healing potential of adipose-derived stem cells in a novel human primary keratinocyte-based scratch assay". International Journal of Molecular Medicine 39.3 (2017): 587-594.
Chicago
Riis, S., Newman, R., Ipek, H., Andersen, J. I., Kuninger, D., Boucher, S., Vemuri, M. C., Pennisi, C. P., Zachar, V., Fink, T."Hypoxia enhances the wound-healing potential of adipose-derived stem cells in a novel human primary keratinocyte-based scratch assay". International Journal of Molecular Medicine 39, no. 3 (2017): 587-594. https://doi.org/10.3892/ijmm.2017.2886
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