A Simplified Protocol for the Purification of Schwann Cells and Exosome Isolation From C57bl/6 Mice



Shojapour M¹ ; Mosayebi G² ; Hajihossein R^{3, 4} ; Noorbakhsh F⁴ ; Mokarizadeh A⁵ ; Ghahremani MH¹ Show Affiliations
Source: Reports of Biochemistry and Molecular Biology Published:2017

Abstract

Background: The purification of Schwann cells has proven to be a difficult process, with most methods requiring the use of special equipment. However, obtaining a sufficient number and high purity of Schwann cells is an integral aspect in their use for clinical application. Therefore, the aim of this study was to establish a simple and effective protocol for the isolation and purification of Schwann cells from the sciatic nerve of C57BL/6 mice. Furthermore, we aimed to provide a protocol for the isolation of exosomes from these cells. Methods: To purify Schwann cells, we used a combination of in situ nerve pre-degeneration and fetal bovine serum. To determine the most effective method of cell purification, we treated the culture with varying concentrations of fetal bovine serum and examined which concentration provided the highest Schwann cell purity. Exosomes were then isolated from Schwann cells through a process of repeated centrifugation and filtration steps. Results: We were able to increase the purified population of Schwann cells from C57BL/6 mice by reducing the concentration of FBS. The purity of Schwann cells at FBS concentrations of 10%, 5%, and 2% were 93.42%, 91.25%, and 97.83%, respectively. Conclusions: When using a concentration of 2% FBS, we obtained the highest purification yield of Schwann cells. Our protocol does not require special equipment or materials. We have created a protocol that is simple, fast, and safe while providing a high yield of purified Schwann cells. The exosome isolation method described in this paper is an appropriate approach with a high quality and yield. © 2018, Varastegan Institute for Medical Sciences.