Neural precursors derived from human embryonic stem cells maintain long-term proliferation without losing the potential to differentiate into all three neural lineages, including dopaminergic neurons
Neural precursors derived from human embryonic stem cells maintain long-term proliferation without losing the potential to differentiate into all three neural lineages, including dopaminergic neurons
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DOI：
10.1111/j.1471-4159.2007.04952.x
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发表时间：
2008-01-01
影响因子：
4.7
通讯作者：
Kim, Kwang-Soo
中科院分区：
文献类型：
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作者：
Hong, Sunghoi;Kang, Un Jung;Kim, Kwang-Soo
人类胚胎干细胞（hES）具有自我更新的能力，并在暴露于适当的信号时分化成多种细胞类型。特别地，hES细胞分化成确定的神经谱系（例如神经元、星形胶质细胞和少突胶质细胞）的能力对于开发用于神经退行性疾病的基于细胞的疗法和研究发育机制是基础。然而，由于缺乏明确的方法来维持其自我更新和指导其分化，hES细胞用于基础和应用研究受到阻碍。最近，我们报道了小鼠ES细胞来源的神经前体细胞（NP）在体外显著扩增后仍具有分化为多巴胺能（DA）神经元的潜能。我们假设来源于hES细胞的NP细胞（hES-NP）也可以进行相同的体外扩增和分化。为了验证这一假设，我们对hES-NP细胞进行了传代培养，并对其增殖和发育特性进行了分析，发现hES-NP细胞在体外扩增12周后可增殖约380000倍，并保持其生成Tuj 1（+）神经元、GFAP（+）星形胶质细胞、O 4（+）少突胶质细胞以及酪氨酸羟化酶阳性（TH（+））DA神经元的潜能。此外，来自hES-NP细胞的TH（+）神经元表达其他中脑DA标记物，包括Nurr 1、Pitx 3、Engrail-1和芳香族L-氨基酸脱羧酶，并释放大量DA。此外，hES-NP细胞通过在液氮中长期储存（超过2年）和多次冻融循环保持其发育潜力。这些结果表明，hES-NP细胞具有提供可发展成特定神经元和神经胶质亚型的可扩展和无限的人细胞来源的能力。
Human embryonic stem (hES) cells have the ability to renew themselves and differentiate into multiple cell types upon exposure to appropriate signals. In particular, the ability of hES cells to differentiate into defined neural lineages, such as neurons, astrocytes, and oligodendrocytes, is fundamental to developing cell-based therapies for neurodegenerative disorders and studying developmental mechanisms. However, the utilization of hES cells for basic and applied research is hampered by the lack of well-defined methods to maintain their self-renewal and direct their differentiation. Recently we reported that neural precursor (NP) cells derived from mouse ES cells maintained their potential to differentiate into dopaminergic (DA) neurons after significant expansion in vitro. We hypothesized that NP cells derived from hES cells (hES-NP) could also undergo the same in vitro expansion and differentiation. To test this hypothesis, we passaged hES-NP cells and analyzed their proliferative and developmental properties.We found that hES-NP cells can proliferate approximately 380 000-fold after in vitro expansion for 12 weeks and maintain their potential to generate Tuj1(+) neurons, GFAP(+) astrocytes, and O4(+) oligodendrocytes as well as tyrosine hydroxylase-positive (TH(+)) DA neurons. Furthermore, TH(+) neurons originating from hES-NP cells expressed other midbrain DA markers, including Nurr1, Pitx3, Engrail-1, and aromatic L-amino acid decarboxylase, and released significant amounts of DA. In addition, hES-NP cells maintained their developmental potential through long-term storage (over 2 years) in liquid nitrogen and multiple freeze-thaw cycles. These results demonstrate that hES-NP cells have the ability to provide an expandable and unlimited human cell source that can develop into specific neuronal and glial subtypes.