Efficient generation of midbrain and hindbrain neurons from mouse embryonic stem cells
Efficient generation of midbrain and hindbrain neurons from mouse embryonic stem cells
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DOI：
10.1038/76536
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发表时间：
2000-06-01
影响因子：
46.9
通讯作者：
McKay, RD
中科院分区：
文献类型：
--
作者：
Lee, SH;Lumelsky, N;McKay, RD
胚胎干细胞是由发育中的胚泡内细胞团分化而来的克隆性细胞系，可在体外广泛增殖，并能适应发育中的胚胎的所有细胞命运。研究表明，从内细胞团1、2或发育中的生殖细胞3中分离出具有ES特性的人类细胞可以提供体细胞前体来源，从而激发了临床对ES细胞的兴趣。以前的研究已经确定了促进特定体细胞命运发育的体外条件，特别是造血细胞、中胚层和神经直肠胚层4、5、6、7。在本研究中，我们提出了一种从小鼠ES细胞体外高产量获得多巴胺(DA)和5-羟色胺能神经元的方法。此外，我们还证明了可以获得无限数量的ES细胞，并且这些类型的神经元是有效生成的。我们从ES细胞中产生CNS祖细胞群体，扩增这些细胞，并在有丝分裂原和特定信号分子的存在下促进它们分化为多巴胺能和5-羟色胺能神经元。神经细胞的分化和成熟是在有丝分裂原从生长介质中退出后完成的。这一实验系统为分析在体外和体内控制这些神经元功能的分子机制提供了强有力的工具，并有可能为理解和治疗神经退行性疾病和精神疾病提供了潜在的工具。
Embryonic stem (ES) cells are clonal cell lines derived from the inner cell mass of the developing blastocyst that can proliferate extensively in vitro and are capable of adopting all the cell fates in a developing embryo. Clinical interest in the use of ES cells has been stimulated by studies showing that isolated human cells with ES properties from the inner cell mass 1, 2 or developing germ cells 3 can provide a source of somatic precursors. Previous studies have defined in vitro conditions for promoting the development of specific somatic fates, specifically, hematopoietic, mesodermal, and neurectodermal 4, 5, 6, 7. In this study, we present a method for obtaining dopaminergic (DA) and serotonergic neurons in high yield from mouse ES cells in vitro. Furthermore, we demonstrate that the ES cells can be obtained in unlimited numbers and that these neuron types are generated efficiently. We generated CNS progenitor populations from ES cells, expanded these cells and promoted their differentation into dopaminergic and serotonergic neurons in the presence of mitogen and specific signaling molecules. The differentation and maturation of neuronal cells was completed after mitogen withdrawal from the growth medium. This experimental system provides a powerful tool for analyzing the molecular mechanisms controlling the functions of these neurons in vitro and in vivo, and potentially for understanding and treating neurodegenerative and psychiatric diseases.