Immunohistological and electrophysiological characterization of Globose basal stem cells

Document Type: Original Article


1 Department of Anatomy, Vardhman Mahavir Medical College & Safdarjung Hospital, New Delhi- 110029, India

2 Department of Physical Medicine and Rehabilitation, Christian Medical College Vellore, India

3 Department of Physical Medicine and Rehabilitation, Christian Medical College, Vellore, India

4 Department of Anatomy, Christian Medical College, Vellore, India


Objective(s): In the past few decades, variety of foetal, embryonic and adult stem and progenitor cells have been tried with conflicting outcome for cell therapy of central nervous system injury and diseases. Cellular characteristics and functional plasticity of Globose basal stem cells (GBCs) residing in the olfactory epithelium of rat olfactory mucosa have not been studied in the past by the neuroscientists due to unavailability of specific markers for GBCs. In the present research, we standardized some techniques to isolate GBCs from rat olfactory epithelium in pure form using a highly selective GBC-III antibody passaged through fluorescence activated cell sorter (FACS). We also characterized these cells immunohistologically using various pluripotent stem cell markers. This work also throws some light on ionic channels present on these stem cells which are responsible for their neuron induction potential.
Materials and methods:Globose basal stem cells were isolated from rat olfactory epithelium using GBC-III antibody and were characterized as multipotent stem cells using various neural progenitor markers. Ionic channels on GBCs were studied with voltage clamping.
Results:GBCs could be isolated in pure (99% purity) form and were found to be stained positive for all neural progenitor cell markers. Voltage gated Na+ channels were completely absent, which proves the unexcitable nature of GBCs. Leaky K+ channels were found to be present on the GBC which was of no significance.
Conclusion: This research work can be helpful in understanding the nature [T1] of these stem cells and utilising them in future as potent candidates for neuro-regenerative therapies.


1. Brewer BJ. Regeneration and proliferation of embryonic and adult rat hippocampal neurons in culture. Exp Neurol 1999; 159:237-247.

2. Schwob JE, Youngentob SL, Mezza RC. Reconzstitution of the rat olfactory epithelium after methyl bromide-induced lesion. J Comp Neurol 1995; 359:15–37.

3.  Viktorov IV, Sukhikh GT. Medico-biological aspects of using stem cells. Vestn Ross Akad Med Nauk 2002; 4:24-30.

4. Iwema CL, Fang H, Kurtz DB, Youngentob SL, Schwob JE. Odorant receptor expression patterns are restored in lesion-recovered rat olfactory epithelium. J Neurosci 2004; 24:356–369.

5. Barnett SC, Riddell JS. Olfactory ensheathing cells (OECs) and the treatment of CNS injury: advantages and possible caveats. J Anat 2004; 204:57-67.

6. Ramon-Cueto A, Valverde F. Olfactory bulb ensheathing glia: a unique cell type with axonal growth-promoting properties. Glia 1995; 14:163– 173.

7. Jang W, Lambropoulos J, Woo JK, Peluso CE, Schwob JE. Maintaining epitheliopoietic potency when culturing olfactory progenitors. Exp Neurol 2008; 214:25-36.

8. Caggiano M, Kauer JS, Hunter DD. Globose basal cells are neuronal progenitors in the olfactory epithelium: a lineage analysis using a replication-incompetent retrovirus. Neuron 1994; 13:339– 352.

9. Goldstein BJ, Schwob JE. Analysis of the globose basal cell compartment in rat olfactory epithelium using GBC-1, a new monoclonal antibody against globose basal cells. J Neurosci 1996; 16:4005–4016.

10. Huard JM, Youngentob SL, Goldstein BJ, Luskin MB, Schwob JE. Adult olfactory epithelium contains multipotent progenitors that give rise to neurons and non-neural cells. J Comp Neurol 1998; 400:469–486.

11. Goldstein BJ, Fang H, Youngentob SL, Schwob JE. Transplantation of multipotent progenitors from the adult olfactory epithelium. Neuroreport 1998; 9: 1611–1617.

12. Harris DT, Rogers I. Umbilical cord blood: a unique source of pluripotent stem cells for regenerative medicine. Curr Stem Cell Res Ther 2007; 2:301-309.

13. Calof AL, Mumm JS,  Rim PC, Shou J. The neuronal stem cell of the olfactory epithelium. J  Neurobiol 1998; 36:190– 205.

14. Carter LA, MacDonald JL, Roskams AJ. Olfactory horizontal basal cells demonstrate a conserved multipotent progenitor phenotype. J Neurosci 2004; 24:5670–5683.

15. Beites CL, Kawauchi S, Crocker CE, Calof AL. Identification and molecular regulation of neural stem cells in the olfactory epithelium. Exp Cell Res 2005; 10:309-316.

16. Youngentob SL, Schwob JE, Saha S, Manglapus G, Jubelt B. Functional consequences following infection of the olfactory system by intranasal infusion of the olfactory bulb line variant (OBLV) of mouse hepatitis strain JHM. Chem Senses 2001; 26:953-963.

17. Freudenberger CB. A comparison of the Wistar albino and the long-evans hybrid strain of the norway rat. Am J Anat 1932; 50:293–349.

18. Schwob JE. Neural regeneration and the peripheral olfactory system.  Anat Rec 2002; 269:33–49.

19. Fricker RA, Carpenter MK, Winkler C, Greco C, Gates MA, Björklund A. Site-specific migration and neuronal differentiation of human neural progenitor cells after transplantation in the adult rat brain. J Neurosci 1999; 19:5990-6005.

20. Guo X, Johe K, Molnar P, Davis H, Hickman J. Characterization of a human fetal spinal cord stem cell line, NSI-566RSC, and its induction to functional motoneurons. J Tissue Eng Regen Med 2010; 4:181-193.

21. Jang W, Kim KP, Schwob JE. Nonintegrin laminin receptor precursor protein is expressed on olfactory stem and progenitor cells. J Comp Neurol 2007; 502:367-381.

21. Paus T, Zijdenbos A, Worsley K, Collins DL, Blumenthal J, Giedd JN, et al. Structural maturation of neural pathways in children and adolescents: In vivo study. Science 1999; 283:1908.

23. Paul F. Developmental (myelogenetic) localisation of the cerebral cortex in the human subject.  Lancet 1901;158:1027.

24. Altman J. Are new neurons formed in the brains of adult mammals? Science 1962; 135:1127–1128.

25. Altman J. Autoradiographic investigation of cell proliferation in the brains of rats and cats. Anat Rec 1963; 145:573–591.

26. Bayer S, Altman J. Hippocampal development in the rat: cytogenesis and morphogenesis examined with autoradiography and low-level X-irradiation. J Comp Neurol 1974; 158:55–79.

27. Giovanna P, Paolo P, Luca B, Thomas RA. Genesis of neuronal and glial progenitors in the cerebellar cortex of peripuberal and adult rabbits. PLoS ONE 2008; 3:2366.

28. Girard SD, Devéze A, Nivet E, Gepner B, Roman FS, Féron F. Isolating nasal olfactory stem cells from rodents or humans. J Vis Exp 2011; 2762.

29. Chen X, Fang H, Schwob JE. Multipotency of purified, transplanted globose basal cells in olfactory epithelium. J Comp Neurol 2004; 469:457-474.

30. Barraud P, He X, Caldwell MA, Franklin RJ. Secreted factors from olfactory mucosa cells expanded as free-floating spheres increase neurogenesis in olfactory bulb neurosphere cultures. BMC Neurosci 2008; 9:24.

31. Krolewski RA, Packard A, Schwob JE. Global expression profiling of globose basal cells and neurogenic progression within the olfactory epithelium. J Comp Neurol 2013; 521:833–859.

32. Gerashchenko BI. Choosing a cell sorting option to study the fate of bystander cells: FACS or MACS? Cytometry 2011; 79A:179–180.