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A Novel Approach for Mucosal and Bulbar Olfactory Ensheathing Cell Isolation Based on the Non-Adherent Subculture Technique

Olfactory ensheathing cells (OECs) are widely used in transplantation studies. Therefore, the high purification of this unique cell type will be valuable for medical applications. Although recent improvements in OEC isolation procedures opened a new era in this field, the high purification efficacy and viability rate are still needed to be considered. The most widely used OECs isolation techniques can be broadly classified as based on adherence properties in particular in the case of olfactory bulb-derived OECs isolation. Considering the invasive nature of harvesting OECs from human olfactory bulbs, hence the high efficient purification of these cells from olfactory mucosa can be beneficial in clinical trials. In this study, we isolated OECs from both the olfactory bulb and mucosa of rats due to their differential adherence properties and compared them. Cell preparations were characterized by NGFR p75 and S100β antibodies, the specific markers for OECs using immunocytochemistry and western blot analysis respectively. OECs morphology and viability were monitored over time by microscopy and MTT assay. We found that utilizing our suggested method, OECs could be purified from the olfactory mucosa as efficiently as the olfactory bulb. Both sources-derived OECs showed high levels of NGFR p75 and S100β expression, although the S100β expression was higher in olfactory mucosa-derived OECs preparations (P<0.05). Moreover, there was no significant difference between the two sources in cell viability in our suggested protocol. Hence due to the non-invasive harvesting method, olfactory mucosa-derived OECs are preferred from a clinical point of view for transplantation studies.