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Cytotechnology

, Volume 68, Issue 4, pp 629–636 | Cite as

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

  • Nazila Niapour
  • Behnam Mohammadi-Ghalehbin
  • Mohammad Ghasem Golmohammadi
  • Mohammad Reza Gholami
  • Mohammad Amani
  • Ali NiapourEmail author
Original Research

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.

Keywords

Schwann cell Rat sciatic nerve KOSR Defined culture 

Notes

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.

Supplementary material

10616_2014_9810_MOESM1_ESM.doc (144 kb)
Supplementary material 1 (DOC 143 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Nazila Niapour
    • 1
  • Behnam Mohammadi-Ghalehbin
    • 2
  • Mohammad Ghasem Golmohammadi
    • 1
  • Mohammad Reza Gholami
    • 3
  • Mohammad Amani
    • 4
  • Ali Niapour
    • 1
    Email author
  1. 1.Department of Anatomical Sciences, School of MedicineArdabil University of Medical SciencesArdabilIran
  2. 2.Department of Microbiology and Parasitology, School of MedicineArdabil University of Medical SciencesArdabilIran
  3. 3.Department of Anatomical Sciences, School of MedicineLorestan University of Medical SciencesKhorramabadIran
  4. 4.Department of Physiology, School of MedicineArdabil University of Medical SciencesArdabilIran

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