Long-term tripotent differentiation capacity of human neural stem (NS) cells in adherent culture
Long-term tripotent differentiation capacity of human neural stem (NS) cells in adherent culture
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DOI：
10.1016/j.mcn.2008.02.014
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发表时间：
2008-06-01
影响因子：
3.5
通讯作者：
Smith, Austin
中科院分区：
文献类型：
--
作者：
Sun, Yirui;Pollard, Steven;Smith, Austin
提供可再生和可扩展的中枢神经系统细胞类型供应的干细胞系将成为基础和应用神经生物学的宝贵资源。在这里，我们描述了在单层培养中多个人类胎儿神经干（NS）细胞系的产生和长期扩增，无需遗传永生化。贴壁的人 NS 细胞在表皮生长因子 (ELF) 和成纤维细胞生长因子 2 (FGF2) 存在的情况下繁殖，在这种条件下，它们稳定表达神经前体标记物，并且向神经元或神经胶质细胞的分化可忽略不计。然而，当它们接触适当的分化因子时，会产生星形胶质细胞、少突胶质细胞和神经元。单细胞克隆证明人类 NS 细胞具有三能性。经过 100 代之后，它们仍保留二倍体核型和恒定的神经发生能力。与人类神经球相比，我们观察到贴壁人类 NS 细胞的持续扩增不需要细胞因子白血病抑制因子 (LIF)。人类 NS 细胞可以稳定转染，以提供报告细胞系，并在活的单层培养物中轻松成像，从而为高内涵遗传和化学筛选创造了潜力。 (C) 2008 Elsevier Inc. 保留所有权利。
Stem cell lines that provide a renewable and scaleable supply of central nervous system cell types would constitute an invaluable resource for basic and applied neurobiology. Here we describe the generation and long-term expansion of multiple human foetal neural stem (NS) cell lines in monolayer culture without genetic immortalization. Adherent human NS cells are propagated in the presence of epidermal growth factor (ELF) and fibroblast growth factor 2 (FGF2), under which conditions they stably express neural precursor markers and exhibit negligible differentiation into neurons or glia. However, they produce astrocytes, oligodendrocytes, and neurons upon exposure to appropriate differentiation factors. Single cell cloning demonstrates that human NS cells are tripotent. They retain a diploid karyotype and constant neurogenic capacity after over 100 generations. In contrast to human neurospheres, we observe no requirement for the cytokine leukaemia inhibitory factor (LIF) for continued expansion of adherent human NS cells. Human NS cells can be stably transfected to provide reporter lines and readily imaged in live monolayer cultures, creating the potential for high content genetic and chemical screens. (C) 2008 Elsevier Inc. All rights reserved.