Othman M, Lu C, Klueber K, Winstead W, Roisen Fj
Department of Anatomical Sciences and Neurobiology, University of Louisville, School of Medicine, Louisville, Kentucky 40292, USA.
Biotech Histochem. 2005 Sep-Dec;80(5-6):189-200. doi: 10.1080/10520290500469777.
Olfactory neuroepithelium (ONe) is unique because it contains progenitor cells capable of mitotic division that replace damaged or lost neurons throughout life. We isolated populations of ONe progenitors from adult cadavers and patients undergoing nasal sinus surgery that were heterogeneous and consisted of neuronal and glial progenitors. Progenitor lines have been obtained from these cultures that continue to divide and form nestin positive neurospheres. In the present study, we used clonal and population analyses to probe the self-renewal and multipotency of the neurosphere forming cells (NSFCs). NSFCs plated at the single cell level produced additional neurospheres; dissociation of these spheres resulted in mitotically active cells that continued to divide and produce spheres as long as they were subcultured. The mitotic activity of clonal NSFCs was assessed using bromodeoxyuridine (BrdU) incorporation. Lineage restriction of the clonal cultures was determined using a variety of antibodies that were characteristic of different levels of neuronal commitment: ss-tubulin isotype III, neural cell adhesion molecule (NCAM) and microtubule associated protein (MAP2), or glial restriction: astrocytes, glial fibrillary acidic protein (GFAP); and oligodendrocytes, galactocerebroside (GalC). Furthermore, nestin expression, a marker indicative of progenitor nature, decreased in defined medium compared to serum-containing medium. Therefore, adult human ONe-derived neural progenitors retain their capacity for self-renewal, can be clonally expanded, and offer multipotent lineage restriction. Therefore, they are a unique source of progenitors for future cell replacement strategies in the treatment of neurotrauma and neurodegenerative diseases.
嗅觉神经上皮（ONe）很独特，因为它含有能够进行有丝分裂的祖细胞，这些祖细胞在整个生命过程中可替代受损或丢失的神经元。我们从成年尸体和接受鼻窦手术的患者中分离出了ONe祖细胞群体，这些群体具有异质性，由神经元祖细胞和神经胶质祖细胞组成。已经从这些培养物中获得了祖细胞系，它们继续分裂并形成巢蛋白阳性神经球。在本研究中，我们使用克隆分析和群体分析来探究形成神经球的细胞（NSFCs）的自我更新能力和多能性。接种在单细胞水平的NSFCs产生了额外的神经球；这些神经球解离后产生了有丝分裂活性细胞，只要进行传代培养，它们就会继续分裂并产生神经球。使用溴脱氧尿苷（BrdU）掺入法评估克隆NSFCs的有丝分裂活性。使用多种抗体来确定克隆培养物的谱系限制，这些抗体是不同水平神经元分化的特征性标志物：β-微管蛋白同种型III、神经细胞黏附分子（NCAM）和微管相关蛋白（MAP2），或者是神经胶质限制标志物：星形胶质细胞、胶质纤维酸性蛋白（GFAP）；以及少突胶质细胞、半乳糖脑苷脂（GalC）。此外，与含血清培养基相比，在限定培养基中，作为祖细胞性质标志物之一的巢蛋白表达降低。因此，成人来源的ONe神经祖细胞保留了自我更新能力，可以进行克隆扩增，并具有多能谱系限制。所以，它们是未来用于治疗神经创伤和神经退行性疾病的细胞替代策略中独特的祖细胞来源。
