Muniswami Durai Murugan, Kanakasabapathy Indirani, Tharion George
Department of Physical Medicine & Rehabilitation, Christian Medical College, Vellore, Tamil Nadu, India.
Department of Anatomy, Christian Medical College, Vellore, Tamil Nadu, India.
Neural Regen Res. 2017 Nov;12(11):1895-1904. doi: 10.4103/1673-5374.219052.
Spinal cord injury (SCI) is a devastating condition with loss of motor and sensory functions below the injury level. Cell based therapies are experimented in pre-clinical studies around the world. Neural stem cells are located intra-cranially in subventricular zone and hippocampus which are highly invasive sources. The olfactory epithelium is a neurogenic tissue where neurogenesis takes place throughout the adult life by a population of stem/progenitor cells. Easily accessible olfactory neuroepithelial stem/progenitor cells are an attractive cell source for transplantation in SCI. Globose basal cells (GBCs) were isolated from rat olfactory epithelium, characterized by flow cytometry and immunohistochemically. These cells were further studied for neurosphere formation and neuronal induction. T10 laminectomy was done to create drop-weight SCI in rats. On the 9 day following SCI, 5 × 10 cells were transplanted into injured rat spinal cord. The outcome of transplantation was assessed by the Basso, Beattie and Bresnahan (BBB) locomotor rating scale, motor evoked potential and histological observation. GBCs expressed neural stem cell markers nestin, SOX2, NCAM and also mesenchymal stem cell markers (CD29, CD54, CD90, CD73, CD105). These cells formed neurosphere, a culture characteristics of NSCs and on induction, differentiated cells expressed neuronal markers βIII tubulin, microtubule-associated protein 2, neuronal nuclei, and neurofilament. GBCs transplanted rats exhibited hindlimb motor recovery as confirmed by BBB score and gastrocnemius muscle electromyography amplitude was increased compared to controls. Green fluorescent protein labelled GBCs survived around the injury epicenter and differentiated into βIII tubulin-immunoreactive neuron-like cells. GBCs could be an alternative to NSCs from an accessible source for autologous neurotransplantation after SCI without ethical issues.
脊髓损伤(SCI)是一种严重的疾病,会导致损伤平面以下的运动和感觉功能丧失。世界各地的临床前研究都在试验基于细胞的疗法。神经干细胞位于颅内的脑室下区和海马体,这些都是高侵袭性的来源。嗅觉上皮是一种神经发生组织,在成年期,一群干细胞/祖细胞会在此持续进行神经发生。易于获取的嗅觉神经上皮干细胞/祖细胞是脊髓损伤移植中一种有吸引力的细胞来源。从大鼠嗅觉上皮中分离出球状基底细胞(GBCs),通过流式细胞术和免疫组织化学进行表征。对这些细胞进一步研究其神经球形成和神经元诱导情况。对大鼠进行T10椎板切除术以造成重物坠落性脊髓损伤。在脊髓损伤后的第9天,将5×10个细胞移植到受伤的大鼠脊髓中。通过Basso、Beattie和Bresnahan(BBB)运动评分量表、运动诱发电位和组织学观察来评估移植结果。GBCs表达神经干细胞标志物巢蛋白、SOX2、神经细胞黏附分子(NCAM),也表达间充质干细胞标志物(CD29、CD54、CD90、CD73、CD105)。这些细胞形成神经球,这是神经干细胞的一种培养特征,在诱导后,分化细胞表达神经元标志物βIII微管蛋白、微管相关蛋白2、神经元细胞核和神经丝。与对照组相比,移植了GBCs的大鼠经BBB评分证实后肢运动功能得到恢复,腓肠肌肌电图幅度增加。绿色荧光蛋白标记的GBCs在损伤中心周围存活并分化为βIII微管蛋白免疫反应性神经元样细胞。GBCs可以作为一种替代来源,从易于获取的自体神经移植中获取神经干细胞,用于脊髓损伤后的移植,且不存在伦理问题。