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Diabetic sera disrupted the normal exosome signaling pathway in human mesenchymal stem cells in vitro

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Abstract

Human mesenchymal stem cells were exposed to diabetic sera for 7 days. Cell viability and apoptosis rate were detected by MTT and flow cytometry assays. The expression of key genes such as CD63, Alix, Rab27a, Rab27b, and Rab8b was monitored by real-time PCR. We also measured acetylcholinesterase activity and size and zeta potential of exosomes in the supernatant form diabetic cells and control. The cellular distribution of CD63 was shown by immunofluorescence imaging and western blotting. Any changes in the ultrastructure of cells were visualized by electron microscopy. Data showed a slight decrease in survival rate and an increased apoptosis in diabetic cells as compared to control (p < 0.05). By exposing cells to diabetic sera, a significant increase in the level of all genes CD63, Alix, Rab27a, Rab27b, and Rab8b was observed (p < 0.05). Flow cytometry analysis and immunofluorescence imaging confirmed increasing CD63 protein content upon treatment with diabetic sera (p < 0.05). We found an enhanced acetylcholinesterase activity in a diabetic condition which coincided with the increasing size of exosomes and decrease in zeta potential (p < 0.05). The fatty acid profile was not significantly affected by diabetic sera. Ultrastructural examination detected more accumulated cytoplasmic lipid vacuoles in diabetic cells.

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Acknowledgments

The authors gratefully thank the personnel of Stem Cell Research Center, Tabriz University of Medical Sciences.

Funding

This work was supported by a grant (no. TBZMED.REC.1394.928) from the Research Council, Tabriz University of Medical Sciences.

Author information

Author notes

  1. Reza Rahbarghazi and Malek Soleimani Mehranjani contributed equally to this work.

Authors and Affiliations

  1. Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

    Jafar Rezaie & Reza Rahbarghazi

  2. Department of Biology, Faculty of Sciences, Urmia University, Urmia, Iran

    Vahid Nejati

  3. Department of Pathology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran

    Majid Khaksar & Ahmad Oryan

  4. Endocrine and Metabolism Section, Department of Internal Medicine, Tabriz University of Medical Sciences, Tabriz, Iran

    Nasser Aghamohamadzadeh

  5. Department of Biology, Faculty of Sciences, Arak University, Arak, 3815688349, Iran

    Mohammad Ali Shariatzadeh & Malek Soleimani Mehranjani

  6. Department of Applied Cell Sciences, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran

    Reza Rahbarghazi

  7. Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

    Reza Rahbarghazi

Authors
  1. Jafar Rezaie
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  2. Vahid Nejati
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  3. Majid Khaksar
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  4. Ahmad Oryan
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  5. Nasser Aghamohamadzadeh
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  6. Mohammad Ali Shariatzadeh
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  7. Reza Rahbarghazi
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  8. Malek Soleimani Mehranjani
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Corresponding authors

Correspondence to Reza Rahbarghazi or Malek Soleimani Mehranjani.

Ethics declarations

All volunteers were asked to fill out an informed consent. All procedures performed through the current experiment, involving human participants, were in accordance with the local ethics committee of Tabriz University of Medical Sciences (Ethical code no. TBZMED.REC.1394.928) and ethical principles of Declaration of Helsinki.

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The authors declare that they have no conflict of interest.

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Rezaie, J., Nejati, V., Khaksar, M. et al. Diabetic sera disrupted the normal exosome signaling pathway in human mesenchymal stem cells in vitro. Cell Tissue Res 374, 555–565 (2018). https://doi.org/10.1007/s00441-018-2895-x

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  • DOI: https://doi.org/10.1007/s00441-018-2895-x

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