Bulletin of Experimental Biology and Medicine

, Volume 164, Issue 4, pp 523–527 | Cite as

Preparation of Human Olfactory Ensheathing Cells for the Therapy of Spinal Cord Injuries

  • А. D. Voronova
  • O. V. Stepanova
  • M. P. Valikhov
  • A. V. Chadin
  • А. S. Dvornikov
  • I. V. Reshetov
  • V. P. Chekhonin

We developed an optimal protocol for preparing and culturing of olfactory ensheathing cells from human olfactory mucosa. Using this protocol, we obtained a culture enriched with human olfactory ensheathing cells. Immunofluorescence analysis by simultaneous expression of GFAP and p75NTR markers showed that the content of ensheathing cells was maximum in passage 3 and 4 cultures (94 and 89.5%, respectively). The developed protocol can be recommended for obtaining autologous preparations of human ensheathing cells for cell therapy of spinal cord injuries.

Key Words

olfactory ensheathing cells from human olfactory mucosa human olfactory mucosa cell therapy spinal cord injuries 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Bryukhovetskii AS. Transplantation of Nerve Cells and Tissue Engineering of the Brain in Nervous Diseases. Moscow, 2003. Russian.Google Scholar
  2. 2.
    Viktorov IV, Savchenko EA, Chekhonin VP. Spontaneous neural differentiation of stem cells in culture of human olfactory epithelium. Bull. Exp. Biol. Med. 2007;144(4):596-601.CrossRefPubMedGoogle Scholar
  3. 3.
    Voronova AD, Stepanova OV, Chadin AV, Resheto IV, Chekhonin VP. The Cell Therapy in Traumatic Spinal Cord Injury. Vestn. Ross. Akad. Med. Nauk. 2016;71(6):420-426. Russian.CrossRefPubMedGoogle Scholar
  4. 2.
    Aoki M, Kishima H, Yoshimura K, Ishihara M, Ueno M, Hata K, Yamashita T, Iwatsuki K, Yoshimine T. Limited functional recovery in rats with complete spinal cord injury after transplantation of whole-layer olfactory mucosa: laboratory investigation. J. Neurosurg. Spine. 2010;12(2):122-130.CrossRefPubMedGoogle Scholar
  5. 5.
    Au E, Richter MW, Vincent AJ, Tetzlaff W, Aebersold R, Sage EH, Roskams AJ. SPARC from olfactory ensheathing cells stimulates Schwann cells to promote neurite outgrowth and enhances spinal cord repair. J. Neurosci. 2007;27(27):7208-7221.CrossRefPubMedGoogle Scholar
  6. 6.
    Barber PC, Lindsay RM. Schwann cells of the olfactory nerves contain glial fibrillary acidic protein and resemble astrocytes. Neuroscience. 1982;7(12):3077-3090.CrossRefPubMedGoogle Scholar
  7. 7.
    Barnett SC, Riddell JS. Olfactory ensheathing cell transplantation as a strategy for spinal cord repair ― what can it achieve? Nat. Clin. Pract. Neurol. 2007;3(3):152-161.CrossRefPubMedGoogle Scholar
  8. 8.
    Basso DM, Beattie MS, Bresnahan JC. A sensitive and reliable locomotor rating scale for open field testing in rats. J. Neurotrauma. 1995;12(1):1-21.CrossRefPubMedGoogle Scholar
  9. 9.
    Centenaro LA, Jaeger Mda C, Ilha J, de Souza MA, Kalil-Gaspar PI, Cunha NB, Marcuzzo S, Achaval M. Olfactory and respiratory lamina propria transplantation after spinal cord transection in rats: effects on functional recovery and axonal regeneration. Brain Res. 2011;1426:54-72.CrossRefPubMedGoogle Scholar
  10. 10.
    De Lorenzo AJ. Electron microscopic observations of the olfactory mucosa and olfactory nerve. J. Biophys. Biochem. Cytol. 1957;3(6):839-850.CrossRefPubMedCentralGoogle Scholar
  11. 11.
    Feron F, Mackay-Sim A. Olfactory ensheathing cells isolated from the lamina propria. Patent WO 2001030982 A1, May 03, 2001.Google Scholar
  12. 12.
    Féron F, Perry C, Cochrane J, Licina P, Nowitzke A, Urquhart S, Geraghty T, Mackay-Sim A. Autologous olfactory ensheathing cell transplantation in human spinal cord injury. Brain. 2005;128(Pt 12):2951-2960.CrossRefPubMedGoogle Scholar
  13. 13.
    Gasser HS. Olfactory nerve fibres. J. Gen. Physiol. 1956;39(4): 473-496.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Higginson JR, Barnett SC. The culture of olfactory ensheathing cells — a distinct glial cell type. Exp. Neurol. 2011;229(1):2-9.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Jarmundowicz W. Methods of the obtaining of olfactory ensheathing cells and their application. Patent WO 2007069927 A2, June 21, 2007.Google Scholar
  16. 16.
    Kocsis JD, Lankford KL, Sasaki M, Radtke C. Unique in vivo properties of olfactory ensheathing cells that may contribute to neural repair and protection following spinal cord injury. Neurosci Lett. 2009;456(3):137-142.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Li J, Lepski G. Cell transplantation for spinal cord injury: a systematic review. Biomed. Res. Int. 2013;2013. ID 786475. doi:
  18. 18.
    Lima C, Pratas-Vital J, Escada P, Hasse-Ferreira A, Capucho C, Peduzzi JD. Olfactory mucosa autografts in human spinal cord injury: a pilot clinical study. J. Spinal Cord Med. 2006;29(3):191-203.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Lindsay SL, Toft A, Griffin J, M M Emraja A, Barnett SC, Riddell JS. Human olfactory mesenchymal stromal cell transplants promote remyelination and earlier improvement in gait co-ordination after spinal cord injury. Glia. 2017;65(4):639-656.Google Scholar
  20. 20.
    Mackay-Sim A, Féron F, Cochrane J, Bassingthwaighte L, Bayliss C, Davies W, Fronek P, Gray C, Kerr G, Licina P, Nowitzke A, Perry C, Silburn PA, Urquhart S, Geraghty T. Autologous olfactory ensheathing cell transplantation in human paraplegia: a 3-year clinical trial. Brain. 2008;131(Pt 9):2376-2386.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Mackay-Sim A, St John JA. Olfactory ensheathing cells from the nose: clinical application in human spinal cord injuries. Exp. Neurol. 2011;229(1):174-180.CrossRefPubMedGoogle Scholar
  22. 22.
    Nori S, Nakamura M, Okano H. Plasticity and regeneration in the injured spinal cord after cell transplantation therapy. Prog. Brain Res. 2017;231:33-56.CrossRefPubMedGoogle Scholar
  23. 23.
    Novikova LN, Lobov S, Wiberg M, Novikov LN. Efficacy of olfactory ensheathing cells to support regeneration after spinal cord injury is influenced by method of culture preparation. Exp. Neurol. 2011;229(1):132-142.CrossRefPubMedGoogle Scholar
  24. 24.
    Richter MW, Fletcher PA, Liu J, Tetzlaff W, Roskams AJ. Lamina propria and olfactory bulb ensheathing cells exhibit differential integration and migration and promote differential axon sprouting in the lesioned spinal cord. J. Neurosci. 2005;25(46):10,700-10,711.CrossRefGoogle Scholar
  25. 25.
    Singh N, Gopal SC, Srivastava RN, Chandra T, Agarwal SP, Singh SK, Gupta DK, Balapure AK. In vitro maintenance of olfactory mucosa with enriched olfactory ensheathing cells. J. Stem Cell Res. Ther. 2013;3:132.CrossRefGoogle Scholar
  26. 26.
    Schwob JE. Neural regeneration and the peripheral olfactory system. Anat. Rec. 2002;269(1):33-49.CrossRefPubMedGoogle Scholar
  27. 27.
    Tabakow P, Jarmundowicz W, Czapiga B, Fortuna W, Miedzybrodzki R, Czyz M, Huber J, Szarek D, Okurowski S, Szewczyk P, Gorski A, Raisman G. Transplantation of autologous olfactory ensheathing cells in complete human spinal cord injury. Cell Transplant. 2013;22(9):1591-1612.CrossRefPubMedGoogle Scholar
  28. 28.
    Toft A, Scott DT, Barnett SC, Riddell JS. Electrophysiological evidence that olfactory cell transplants improve function after spinal cord injury. Brain. 2007;130(Pt 4):970-984.PubMedGoogle Scholar
  29. 29.
    Zadroga A, Jezierska-Woźniak K, Czarzasta J, Barczewska M, Wojtkiewicz J, Maksymowicz W. Therapeutic potential of olfactory ensheathing cells and mesenchymal stem cells in spinal cord injurie. Stem Cells Int. 2017;2017. ID 3978595. doi:
  30. 30.
    Zhang SX, Huang F, Gates M, White J, Holmberg EG. Histological repair of damaged spinal cord tissue from chronic contusion injury of rat: a LM observation. Histol. Histopathol. 2011;26(1):45-58.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • А. D. Voronova
    • 1
    • 2
  • O. V. Stepanova
    • 2
  • M. P. Valikhov
    • 2
  • A. V. Chadin
    • 2
  • А. S. Dvornikov
    • 2
  • I. V. Reshetov
    • 3
  • V. P. Chekhonin
    • 1
    • 2
  1. 1.Department of Fundamental and Applied Neurobiology, V. P. Serbsky Federal Medical Research Centre for Psychiatry and NarcologyMinistry of Health of the Russian FederationMoscowRussia
  2. 2.Department of Medical Nanobiotechnologies, Medical and Biological FacultyN. I. Pirogov National Research Medical UniversityMoscowRussia
  3. 3.University Clinical Hospital No. 1, I. M. Sechenov First Moscow State Medical UniversityMinistry of Health of the Russian FederationMoscowRussia

Personalised recommendations