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基于非贴壁传代培养技术的黏膜和球部嗅鞘细胞分离新方法。

A Novel Approach for Mucosal and Bulbar Olfactory Ensheathing Cells Isolation Based on the Non-adherent Subculture Technique.

作者信息

Tirgar Fatemeh, Azizi Zahra, Hadjighassem Mahmoudreza

机构信息

Department of Neuroscience and Addiction Studies, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.

Department of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.

出版信息

Basic Clin Neurosci. 2024 Mar-Apr;15(2):211-220. doi: 10.32598/bcn.2022.3579.1. Epub 2024 Mar 1.

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.

摘要

引言

嗅鞘细胞(OECs)广泛应用于移植研究。这种独特细胞类型的高度纯化对于医学应用具有重要价值。尽管最近嗅鞘细胞分离程序的改进开启了该领域的新纪元,但高纯化效率和存活率仍是人们关注的问题。最广泛使用的嗅鞘细胞分离技术可根据贴壁特性大致分类,特别是在源自嗅球的嗅鞘细胞分离方面。考虑到从人嗅球采集嗅鞘细胞具有侵入性,从嗅黏膜高效纯化这些细胞将有利于临床试验。在本研究中,我们基于大鼠嗅球和黏膜不同的贴壁特性分离了嗅鞘细胞并进行比较。

方法

分别使用免疫细胞化学和蛋白质印迹分析,通过OECs的特异性标志物NGFR p75和S100β抗体对细胞制剂进行表征。通过显微镜检查和MTT(3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐)试验随时间监测嗅鞘细胞的形态和活力。

结果

我们发现,使用我们建议的方法可以从嗅黏膜中高效纯化嗅鞘细胞,其效率与从嗅球中纯化的相当。两种来源的嗅鞘细胞均显示出高水平的NGFR p75和S100β表达,尽管源自嗅黏膜的嗅鞘细胞制剂中S100β表达更高(P<0.05)。此外,在我们建议的方案中,两种来源的细胞活力没有显著差异。

结论

由于采用非侵入性采集方法,从临床角度来看,源自嗅黏膜的嗅鞘细胞在移植研究中更受青睐。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9907/11367208/1186fb37f619/BCN-15-211-g001.jpg

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