Thakur Avinash, Muniswami Duraimurugan, Tharion George, Kanakasabapathy Indirani
Assistant Professor, Department of Anatomy, Vardhman Mahavir Medical College & Safdarjung Hospital , New Delhi- 110029, India .
Faculty, Department of Physical Medicine and Rehabilitation, Christian Medical College Vellore, India .
J Clin Diagn Res. 2013 Nov;7(11):2419-22. doi: 10.7860/JCDR/2013/6372.3562. Epub 2013 Nov 10.
Olfactory mucosa which is situated in the roof of the nasal cavity possesses an extremely peculiar and exceptional type of pluripotent stem cells called Globose Basal Cells (GBCs) which help in lifelong regeneration of the olfactory mucosa. Previous literature doesn't provide much knowledge on the cytological, histochemical and electrophysiological properties of these cells, as they have never been isolated in pure form.
Olfactory mucosa was obtained from six Albino Wistar rats by using standardized surgical and chemical separation procedures. GBCs were isolated by using different chemical, surgical and fluorescent techniques.
In this research work, we standardized the techniques for isolating these stem cells in pure form from rat olfactory mucosa by tagging them with GBC-III antibody and separating them from other epithelial cells by using Fluorescence Activated Cell Sorter (FACS). GBC-III antibody is a mouse monoclonal IgM antibody which recognizes a 40 kDa surface antigen, which is a laminin receptor surface protein present on the GBCs. It is a highly specific marker for GBCs, unlike the earlier antibodies used, like GBC-I, which were nonspecific markers for GBCs and showed positive reactions, even with Horizontal Basal Cells (HBCs), sustentacular cells (Sus) and duct cells. This study also standardized the techniques for surgically excising the olfactory mucosa from the nasal septum and chemically separating the olfactory epithelium from the lamina propria.
GBCs are an important group of cells which can be exploited in future to study and treat neuro-degenerative disorders like multiple sclerosis, brain ischaemia, etc. and spinal cord trauma, as they reside in a niche similar to the microenvironment in the central nervous system and have the similar ectodermal development as the neuronal and non-neuronal cells of the CNS. Moreover, olfactory epithelium is easily accessible for autologous transplantation of GBCs for different CNS disorders.
位于鼻腔顶部的嗅黏膜拥有一种极为特殊的多能干细胞,称为球状基底细胞(GBCs),它有助于嗅黏膜的终身再生。以往文献对这些细胞的细胞学、组织化学和电生理特性了解甚少,因为它们从未以纯形式分离出来。
通过标准化的手术和化学分离程序,从六只白化Wistar大鼠获取嗅黏膜。使用不同的化学、手术和荧光技术分离GBCs。
在本研究工作中,我们通过用GBC-III抗体标记并使用荧光激活细胞分选仪(FACS)将它们与其他上皮细胞分离,从而标准化了从大鼠嗅黏膜中以纯形式分离这些干细胞的技术。GBC-III抗体是一种小鼠单克隆IgM抗体,可识别一种40 kDa的表面抗原,该抗原是GBCs上存在的层粘连蛋白受体表面蛋白。它是GBCs的高度特异性标志物,与早期使用的抗体如GBC-I不同,GBC-I是GBCs的非特异性标志物,甚至对水平基底细胞(HBCs)、支持细胞(Sus)和导管细胞也呈阳性反应。本研究还标准化了从鼻中隔手术切除嗅黏膜以及从固有层化学分离嗅上皮的技术。
GBCs是一类重要的细胞,未来可用于研究和治疗神经退行性疾病,如多发性硬化症、脑缺血等以及脊髓损伤,因为它们所处的微环境与中枢神经系统相似,并且与中枢神经系统的神经元和非神经元细胞具有相似的外胚层发育。此外,嗅上皮易于获取,可用于针对不同中枢神经系统疾病进行GBCs的自体移植。