Efficacy of optimized in vitro predegeneration period on the cell count and purity of canine Schwann cell cultures


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

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

3 Department of Physiology, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran

4 Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran


Objective(s): Predegeneration is a standard technique to obtain mitotically activated and enriched cultures of Schwann cells (SCs). This study, for the first time, evaluated the impact of various duration of predegeneration on cell yield and enrichment of SCs from dog peripheral nerve.
Materials and Methods: Dog sural nerves were subjected to 5, 10, 15 day-long in vitro predegeneration. The total cell yield and the purity of SCs were evaluated in each group on the first and seventh day after plating.
Results: The maximum and minimum numbers of cells were counted in 15 day-long predegene-ration and control groups which underwent no predegeneration. The 10 day-long in vitro predegeneration group with 80±0.5% SCs enrichment had the best purity after plating day and could maintain its purity with elapsing on cultures.
Conclusion: 10 day-long predegeneration results in the higher cell number and the better and prolonged purity of SCs in culture.


1. Kocsis JD, Waxman SG. Schwann cells and their precursors for repair of central nervous system myelin. Brain 2007; 130:1978-1980.
2. Perin EC, Silva GV, Assad JA, Vela D, Buja LM, Sousa AL, et al. Comparison of intracoronary and transendocardial delivery of allogeneic mesenchymal cells in a canine model of acute myocardial infarction. J Mol Cell Cardilo 2008; 44:486-495.
3. Niapour A, Karamali F, Karbalaie K, Kiani A, Mardani M, Nasr-Esfahani MH, et al. Novel method to obtain highly enriched cultures of adult rat Schwann cells. Biotechnol Lett 2010; 32:781-786.
4. Gołka B, Lewin-Kowalik J, S´wie¸ch-Sabuda E, Larysz-Brysz M, Go´rka D, Małecka-Tendera E. Predegenerated peripheral nerve grafts rescue retinal ganglion cells from axotomy-induced death. Exp Neurol 2001; 167:118-125.
5. Kraus A, Tager J, Kohler K, Manoli T, Haerle M, Werdin F, et al. Efficacy of various durations of in vitro predegeneration on the cell count and purity of rat Schwann-cell cultures. J Neurotrauma 2010; 27:197-203.
6. Catchpole B, Ristic JM, Fleeman LM, Davison LJ. Canine diabetes mellitus: can old dogs teach us new tricks? Diabetologia 2005; 48:1948-1956.
7. Gandolfi F, Vanelli A, Pennarossa G, Rahaman M, Acocella F, Brevini TA. Large animal models for cardiac stem cell therapies. Theriogenology 2011; 75:1416-1425.
8. Harding J, Roberts RM, Mirochnitchenko O. Large animal models for stem cell therapy. Stem Cell Res Ther 2013; 4:23.
9. Haastert K, Seef P, Stein VM, Tipold A, Grothe C. A new cell culture protocol for enrichment and genetic modification of adult canine Schwann cells suitable for peripheral nerve tissue engineering. Res Vete Sci 2009; 87:140-142.
10. de la Fuente I, Alcalde I, Gamboa OL, Garrosa M,
Gayoso MJ. A method for obtaining Schwann cell cultures from adult rabbit nerve based on "in vitro" pre-degeneration and neuregulin treatment. Histol Histopathol 2012; 27:95-102.
11. Liu J, Wang J, Luo F, Wang Z, Wang Y. Reconstruction of the abdominal vagus nerve using sural nerve grafts in canine models. PloS One 2013; 8:e58903.
12. David S, Aguayo AJ. Axonal elongation into peripheral nervous system ‘bridges’ after central nervous system injury in adult rats. Science 1981; 214:931-393.
13. Keilhoff G, Fansa H, Schneider W, Wolf G. In vivo predegeneration of peripheral nerves: an effective technique to obtain activated Schwann cells for nerve conduits. J Neurosci Methods 1999; 89:17-24.
14. Dai LG, Huang GS, Hsu SH. Sciatic nerve regeneration by cocultured Schwann cells and stem cells on microporous nerve conduits. Cell Transplant 2013; 22:2029-2039.
15. Hill CE, Brodak DM, Bartlett Bunge M. Dissociated predegenerated peripheral nerve transplants for spinal cord injury repair: a comprehensive assessment of their effects on regeneration and functional recovery compared to Schwann cell transplants. J Neurotrauma 2012; 29:2226-2243.
16. Zhelyaznik N, Mey J. Regulation of retinoic acid receptors α, β and retinoid X receptor α after sciatic nerve injury. Neurosci 2006; 141:1761-1774.
17. Zhang C, Kong X, Ning G, Liang Z, Qu T, Chen F, et al. All-trans retinoic acid prevents epidural fibrosis through NF-kappaB signaling pathway in post-laminectomy rats. Neuropharmacology 2013; 79C:275-281.