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Oxidative Stress Alters the Morphological Responses of Myoblasts to Single-Site Membrane Photoporation

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Abstract

The responses of single cells to plasma membrane damage is critical to cell survival under adverse conditions and to many transfection protocols in genetic engineering. While the post-damage molecular responses have been much studied, the holistic morphological changes of damaged cells have received less attention. Here we document the post-damage morphological changes of the C2C12 myoblast cell bodies and nuclei after femtosecond laser photoporation targeted at the plasma membrane. One adverse environmental condition, namely oxidative stress, was also studied to investigate whether external environmental threats could affect the cellular responses to plasma membrane damage. The 3D characteristics data showed that in normal conditions, the cell bodies underwent significant shrinkage after single-site laser photoporation on the plasma membrane. However for the cells bearing hydrogen peroxide oxidative stress beforehand, the cell bodies showed significant swelling after laser photoporation. The post-damage morphological changes of single cells were more obvious after chronic oxidative exposure than that after acute ones. Interestingly, in both conditions, the 2D projection of nucleus apparently shrank after laser photoporation and distanced itself from the damage site. Our results suggest that the cells may experience significant multi-dimensional biophysical changes after single-site plasma membrane damage. These post-damage responses could be dramatically affected by oxidative stress.

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Abbreviations

DM:

Dichroic mirror

EOM:

Electro-optic modulator

Fs:

Femtosecond

NA:

Numerical aperture

NCD:

The distance between the nucleus center and the membrane damage site

PP:

Photoporation

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Acknowledgments

The authors thank the Hong Kong Research Grants Council for the support of this research project (CUHK 415413) through its General Research Funds.

Conflict of interests

Xinxing Duan, Jennifer MF Wan and Arthur FT Mak declare that they have no conflicts of interest.

Ethical Standards

No human studies were carried out by the authors for this article. No animal studies were carried out by the authors for this article.

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Authors and Affiliations

  1. Division of Biomedical Engineering, The Chinese University of Hong Kong, Shatin, Hong Kong

    Xinxing Duan & Arthur F. T. Mak

  2. Department of Electronic Engineering, The Chinese University of Hong Kong, Shatin, Hong Kong

    Arthur F. T. Mak

  3. Department of Mechanical & Automation Engineering, The Chinese University of Hong Kong, Shatin, Hong Kong

    Xinxing Duan & Arthur F. T. Mak

  4. School of Biological Sciences, The University of Hong Kong, Pokfulam Road, Hong Kong

    Xinxing Duan & Jennifer M. F. Wan

Authors
  1. Xinxing Duan
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  2. Jennifer M. F. Wan
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Correspondence to Arthur F. T. Mak.

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Associate Editor Richard Dickinson oversaw the review of this article.

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Duan, X., Wan, J.M.F. & Mak, A.F.T. Oxidative Stress Alters the Morphological Responses of Myoblasts to Single-Site Membrane Photoporation. Cel. Mol. Bioeng. 10, 313–325 (2017). https://doi.org/10.1007/s12195-017-0488-5

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  • DOI: https://doi.org/10.1007/s12195-017-0488-5

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