Yue Feng, Chen Biao, Wu Di, Dong Kai, Zeng Si-en, Zhang Yu
Department of Neurology, Beijing Institute of Geriatrics, Xuanwu Hospital, Capital University of Medical Science, and Key Laboratory of Neurodegeneration, Ministry of Education, Beijing 100053, China.
Chin Med J (Engl). 2006 Jan 20;119(2):110-6.
The existence of neurogenesis in the hippocampus of adult nonhuman primates has been confirmed in recent years, however, the biological properties of adult neural stem cells or neural progenitor cells (NPCs) from this region remain to be extensively explored. The present work was to investigate on the expansion of NSCs/NPCs from the hippocampus of adult cynomolgus monkeys and the examination of their characteristics in vitro.
NPCs isolated from the hippocampus of adult cynomolgus monkeys were expanded in vitro in serum-free media containing growth factors, and were then allowed to differentiate by removing mitotic factors. The expansion capacity of NPCs and their differentiation potential were assayed by immunohistochemical and immunocytochemical analysis.
During primary culture, NPCs underwent cell division, proliferation and aggregation to form neurospheres that were growing in suspension. Without mitotic stimulation, most neurospheres adhered to the culture dish and started to differentiate. Eventually, nearly 12% of the differentiated cells expressed neuron specific marker-beta III-tubulin (Tuj1) and 84% expressed astrocyte specific marker-fibrillary acidic protein (GFAP). In addition, the expression of a neural stem cell marker, nestin, was found both in NPCs and in the subgranular zone of adult monkey hippocampus, where NPCs were originally derived.
NPCs from the hippocampus of adult cynomolgus monkeys can be expanded to some extent in vitro and are capable of differentiating into neurons and astrocytes. Further experiments to promote the in vitro proliferation capacity of NPCs will be required before adult NPCs can be used as a useful cell model for studying adult neurogenesis and cell replacement therapy using adult stem cells.
近年来已证实成年非人灵长类动物海马体中存在神经发生现象,然而,该区域成年神经干细胞或神经祖细胞(NPCs)的生物学特性仍有待深入探究。本研究旨在探讨成年食蟹猴海马体中神经干细胞/神经祖细胞的体外扩增及其特性检测。
从成年食蟹猴海马体中分离出的NPCs在含有生长因子的无血清培养基中进行体外扩增,然后去除有丝分裂因子使其分化。通过免疫组织化学和免疫细胞化学分析检测NPCs的扩增能力及其分化潜能。
原代培养期间,NPCs进行细胞分裂、增殖并聚集形成悬浮生长的神经球。在没有有丝分裂刺激的情况下,大多数神经球附着于培养皿并开始分化。最终,近12%的分化细胞表达神经元特异性标志物βIII-微管蛋白(Tuj1),84%表达星形胶质细胞特异性标志物纤维酸性蛋白(GFAP)。此外,在NPCs以及成年猴海马体颗粒下区(NPCs的原始来源地)均发现了神经干细胞标志物巢蛋白的表达。
成年食蟹猴海马体中的NPCs能够在体外一定程度上扩增,并能够分化为神经元和星形胶质细胞。在成年NPCs可作为研究成年神经发生和使用成体干细胞进行细胞替代治疗的有用细胞模型之前,还需要进一步实验来提高NPCs的体外增殖能力。