Smith R, Bagga V, Fricker-Gates R A
School of Biosciences, Cardiff University, Cardiff CF10 3US, Wales, UK.
J Hematother Stem Cell Res. 2003 Dec;12(6):713-25. doi: 10.1089/15258160360732731.
One of the major obstacles to the use of neural stem/progenitor cells in neuronal replacement therapy is the limited ability of these cells to generate sufficient numbers of specific neuronal phenotypes either in the culture dish or after transplantation in animal models of neurodegenerative disease. It is not yet fully understood whether embryonic neural stem and progenitor cells show species-specific or regional identities, or if current culture paradigms select for a particular subset of stem cells/progenitors with similar proliferation and differentiation capacities. To investigate this issue, we isolated embryonic neural progenitors derived from the developing rat and mouse central nervous system for in vitro culture to assess the regional, species-specific, and temporal effects on both cell proliferation and generation of neurons. Neurosphere cultures were derived from E13-15 mouse or rat developing striatum (medial, lateral, or whole ganglionic eminence), ventral mesencephalon, and cortex. We compared basic fibroblast growth factor and epidermal growth factor for their influence on cell proliferation and neuronal differentiation under defined differentiation paradigms. Seeding density and conditioned media were also tested for their effects on maintenance of cell proliferation over protracted time periods. Results showed that embryonic neural stem/progenitor cells maintained defined patterns of proliferation and neuronal differentiation, with both declining with time in vitro. Proliferation rate was more dependent on species and region than the neurotrophins or conditions used for culture. These results suggest that the appropriate selection of embryonic neural stem cells and culture conditions may be crucial for the optimization of their neurogenic potential.
在神经元替代疗法中使用神经干细胞/祖细胞的主要障碍之一是，这些细胞在培养皿中或在神经退行性疾病动物模型中移植后，产生足够数量特定神经元表型的能力有限。目前尚不完全清楚胚胎神经干细胞和祖细胞是否表现出物种特异性或区域特性，或者当前的培养模式是否选择了具有相似增殖和分化能力的特定干细胞/祖细胞亚群。为了研究这个问题，我们分离了来自发育中的大鼠和小鼠中枢神经系统的胚胎神经祖细胞进行体外培养，以评估区域、物种特异性和时间对细胞增殖和神经元生成的影响。神经球培养物来源于E13 - 15小鼠或大鼠发育中的纹状体（内侧、外侧或整个神经节隆起）、腹侧中脑和皮质。我们比较了碱性成纤维细胞生长因子和表皮生长因子在特定分化模式下对细胞增殖和神经元分化的影响。还测试了接种密度和条件培养基对长时间维持细胞增殖的影响。结果表明，胚胎神经干细胞/祖细胞维持了特定的增殖和神经元分化模式，两者在体外培养时均随时间下降。增殖率更多地取决于物种和区域，而不是所使用的神经营养因子或培养条件。这些结果表明，胚胎神经干细胞和培养条件的适当选择对于优化其神经发生潜能可能至关重要。
