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Elevated expression of stemness genes in adipose-derived mesenchymal stem cells cultured on fibrin scaffold

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Abstract

In regenerative medicine, MSCs need to be pluripotent for better results. In this study, the effect of fibrin scaffold on expression of stemness genes was examined. Adipose-derived MSCs were cultured in tissue culture plates (2D) and 3-dimensional (3D) fibrin scaffolds. The effect of fibrin scaffold on proliferation of adipose-derived MSCs was evaluated by MTT assay. The expression of stemness genes (OCT4 and SOX2) were evaluated by qRT-PCR, and flow cytometry was done for Nanog protein level. Cultured MSCs on fibrin scaffold were able to proliferate according to data obtained by MTT assay. Expression of OCT4 and SOX2 had a significant increase in cells were cultured in 3D condition compared to 2D condition (P < 0.05). Also, increased expression of Nanog protein in 3D culture was observed (P < 0.05). OCT4 and SOX2 in 3D condition increased two-fold and three-fold respectively in 2D and 3D conditions. Moreover, expression of Nanog increased 30% more than in 2D condition. Evaluation of important pluripotency regulators such as OCT4, SOX2, and Nanog showed that fibrin scaffolds are useful instruments to maintain stemness of MSCs, which is essential in field of stem cell therapy and regenerative medicine.

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Abbreviations

MSC:

mesenchymal stem cell

2D:

two-dimensional culture

3D:

three-dimensional culture

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Acknowledgements

The present study was supported by the Research Deputy of Shahroud University of Medical Sciences. Financial support was provided by Najafabad Branch, Islamic Azad University.

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Author notes

  1. Azadeh Anbarlou

    Present address: Department of Laboratory Medicine, Institute of Biomedicine, Sahlgrenska Academy at University of Gothenburg, Göteborg, Sweden

Authors and Affiliations

  1. Department of Tissue Engineering, Najafabad Branch, Islamic Azad University, Najafabad, Iran

    Arezoo Mousavifard

  2. Department of Biochemistry, Najafabad Branch, Islamic Azad University, Najafabad, Iran

    Elahe Pourazizi Najafabadi

  3. Department of Hematology and Cell Therapy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

    Mahshid Akhavan Rahnama

  4. Department of Tissue Engineering, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Azadeh Anbarlou

  5. Stem Cell and Tissue Engineering Research Center, Shahroud University of Medical Sciences, Shahroud, Iran

    Amir Atashi

Authors
  1. Arezoo Mousavifard
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  2. Elahe Pourazizi Najafabadi
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  3. Mahshid Akhavan Rahnama
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  4. Azadeh Anbarlou
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Corresponding author

Correspondence to Amir Atashi.

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Ethical approval and consent to participate

This experimental study was approved by the Ethical Committee of Islamic Azad University (No. 135896). Adipose tissue of abdomen was obtained from donors undergoing liposuction with the informed consent. Harvested tissues were placed immediately in phosphate-buffered Saline (PBS) (Invitrogen) containing 1% penicillin/streptomycin (Invitrogen). Adipose-derived MSCs were isolated by collagenase digestion method (collagenase type I, Invitrogen). Then, the cells were re-suspended in Dulbecco’s Modified Eagle’s Medium (DMEM, Bio-Idea, Iran) supplemented with 10% Fetal Bovine Serum (FBS) (Invitrogen) and 1% penicillin/streptomycin (Invitrogen) and incubated at 37°C.

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Corresponding editor: BJ Rao

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Mousavifard, A., Pourazizi Najafabadi, E., Akhavan Rahnama, M. et al. Elevated expression of stemness genes in adipose-derived mesenchymal stem cells cultured on fibrin scaffold. J Biosci 45, 81 (2020). https://doi.org/10.1007/s12038-020-00050-5

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  • DOI: https://doi.org/10.1007/s12038-020-00050-5

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