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An efficient system for selection and culture of Schwann cells from adult rat peripheral nerves

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Abstract

Schwann cells (SCs), the supporting cells of the peripheral nerves, are indispensable for regenerating the peripheral and central nervous system. Copious preparation of these cells in a well-defined manner is to be a privileged position. SCs cultivation is overwhelmed by contaminating fibroblasts which are often outgrowing as the predominant cell type in an in vitro culture. This study introduces a technically simple and efficient procedure for SCs isolation and enrichment based on implementing recombinant and defined supplements. Collected adult rat sciatic nerves were cultured for 10 days as in vitro predegeneration. After dissociation and plating, the medium changed to knockout serum replacement supplemented DMDM/F12 medium containing various growth factors. The whole procedure took 3 weeks and SCs purity was then evaluated through implementing specific cytoplasmic and membranous markers. The viability of enriched SCs were evaluated by MTT assay. Within 10 days, over 99 % homogenous SCs were achieved and confirmed through immunofluorescence staining and flow-cytometry for P75NTR and S100 markers, respectively. MTT data revealed that the viability and metabolic activities of purified SCs were increased in expansion medium. This study provides a technically easy and efficient method with the benefits of not utilizing bovine serum or other animal products for SCs isolation and enrichment.

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Acknowledgments

This study was financially supported (Grant No. 91393) by the Vice Chancellor for Research of the Ardabil University of Medical Sciences, Ardabil, Iran.

Conflict of interest

The authors have declared that no competing interests exist.

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Authors and Affiliations

  1. Department of Anatomical Sciences, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran

    Nazila Niapour, Mohammad Ghasem Golmohammadi & Ali Niapour

  2. Department of Microbiology and Parasitology, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran

    Behnam Mohammadi-Ghalehbin

  3. Department of Anatomical Sciences, School of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran

    Mohammad Reza Gholami

  4. Department of Physiology, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran

    Mohammad Amani

Authors
  1. Nazila Niapour
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  2. Behnam Mohammadi-Ghalehbin
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  3. Mohammad Ghasem Golmohammadi
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  4. Mohammad Reza Gholami
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  5. Mohammad Amani
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  6. Ali Niapour
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Correspondence to Ali Niapour.

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Niapour, N., Mohammadi-Ghalehbin, B., Golmohammadi, M.G. et al. An efficient system for selection and culture of Schwann cells from adult rat peripheral nerves. Cytotechnology 68, 629–636 (2016). https://doi.org/10.1007/s10616-014-9810-4

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  • DOI: https://doi.org/10.1007/s10616-014-9810-4

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