Abstract
The human olfactory mucosa is located in the middle and superior turbinates, and the septum of nasal cavity. Olfactory mucosa plays an important role in detection of odours and it is also the only nervous tissue that is exposed to the external environment. This property leads to easy access to the olfactory mucosa for achieving various researches. The lamina propria of olfactory mucosa consists of olfactory ensheathing cells (OECs) that cover the nerve fibers of olfactory. Here we describe a protocol for isolation of OECs from biopsy of human olfactory mucosa.
Keywords: Human olfactory mucosa, Olfactory ensheathing cells (OECs), S100-beta antigen, Primary cell culture
Background
Olfactory ensheathing cells (OECs) are glial cells that express various antigens similar to astrocytes and Schwann cells such as glial fibrillary-associated protein (GFAP), S100-beta, p75 low-affinity nerve growth factor receptor, vimentin, nestin, and neuropeptide Y (Singh et al., 2013). Olfactory ensheathing cells release different neurotrophic factors and adhesion molecules that function in cellular growth and adhesions of central nervous system (Pastrana et al., 2007). In addition, these cells play an important role in the regeneration of the damaged central nervous system such as treatment of spinal cord injury and neurodegenerative diseases (Novikova et al., 2011). We select OECs as research material in our study as they have several advantage properties such as high migratory capacity, accessible source, differentiation from stem cells of nasal olfactory mucosa, and non-tumorigenicity behavior (Huang et al., 2008; Escada et al., 2009). This protocol describes a step-by-step procedure for the isolation of OECs from Human Olfactory Mucosa Specimen.
Materials and Reagents
Equipment
Procedure
Data analysis
Confirmation of cultured primary OECs was done by detecting S100-beta antigen via fluorescence immunocytochemical analysis. This analysis demonstrated that cells expressed S100-beta antigen. Test results are obtained from three independent experiments.
Notes
Recipes
Acknowledgments
This study was supported by grant number 20035 obtained from School of Advanced Technologies in Medicine and grant 19925 from Brain and Spinal Cord injury Research Center, Tehran University of Medical Sciences. The protocol was adapted from Hashemi et al. (2016).
References
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