Tirgar F, Azizi Z, Hadjighassem M. A Novel Approach for Mucosal and Bulbar Olfactory Ensheathing Cells Isolation Based on the Non-adherent Subculture Technique. BCN 2024; 15 (2) :211-220
URL:
http://bcn.iums.ac.ir/article-1-2390-en.html
1- Department of Neuroscience and Addiction Studies, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.
2- Department of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Abstract:
Introduction: Olfactory ensheathing cells (OECs) are widely used in transplantation studies. The high purification of this unique cell type is valuable for medical applications. Although recent improvements in OECs isolation procedures opened a new era in this field, the high purification efficacy and viability rate are still of concern. The most widely used OECs isolation techniques can be broadly classified based on adherence properties, particularly in olfactory bulb-derived OEC isolation. Considering the invasive nature of harvesting OECs from human olfactory bulbs, a highly efficient purification of these cells from olfactory mucosa can benefit clinical trials. In this study, we isolated OECs from rats’ olfactory bulbs and mucosa due to their differential adherence properties and compared them.
Methods: 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 (3-[4,5-dimethylthiazol2-yl]-2,5-diphenyltetrazolium bromide) assay.
Results: We found that OECs could be purified from the olfactory mucosa using our suggested method as efficiently as the olfactory bulb. Both 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.
Conclusion: Due to the non-invasive harvesting method, olfactory mucosa-derived OECs are preferred from a clinical point of view in transplantation studies.
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• Olfactory ensheathing cells (OECs) can be isolated from the olfactory bulb and mucosa for transplantation studies.
• Both derived OEC preparations showed high levels of specific markers.
• Both derived OEC preparations showed similar cell viability.
• OM-derived OECs are preferred due to the non-invasive isolation method.
Plain Language Summary
The olfactory ensheathing cells (OECs) which surround olfactory neurons are utilized in clinical trials to treat many diseases such as brain tumors, stroke, spinal cord injury, and neurorehabilitation. Due to their special properties, such as high migration ability towards damaged areas, these cells can be used as carriers for transferring genes and therapeutic agents to the desired areas of the nervous system. In addition, OECs are involved in repairing damaged tissues and disease recovery by secreting growth factors and anti-inflammatory factors. Since these cells are widely used in the clinic, their isolation and purification are essential issues. OECs can be isolated from two main sources: The olfactory bulb and the olfactory mucosa. Unlike isolation from the olfactory bulb located inside the skull, isolation from the olfactory mucosa within the roof of the nasal cavity is a non-invasive process. Therefore, from a clinical perspective, it is important to find novel and easy strategies to isolate and purify OECs from the olfactory mucosa. In this research, we presented a new method to isolate these cells from the olfactory mucosa as efficiently as the olfactory bulb discussed in previous studies. The cell purity and cell viability of OECs isolated from both sources were compared. The results showed that utilizing our strategy, the OECs derived from the olfactory mucosa have the same properties and viability as the OECs derived from the olfactory bulb. The results of the present study indicate the effectiveness of the suggested process for OEC isolation from olfactory mucosa. This process is non-invasive and can be easily extracted from the roof of the patient`s nasal cavity. It is expected that OECs will be used for cell therapy treatments in the clinic in the near future.
Type of Study:
Original |
Subject:
Cellular and molecular Neuroscience Received: 2021/01/16 | Accepted: 2022/04/12 | Published: 2024/03/1