Neural differentiation of embryonic stem cells in vitro: a road map to neurogenesis in the embryo.
Neural differentiation of embryonic stem cells in vitro: a road map to neurogenesis in the embryo.
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DOI：
10.1371/journal.pone.0006286
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发表时间：
2009-07-21
期刊：
影响因子：
3.7
通讯作者：
Bekman E
中科院分区：
文献类型：
--
作者：
Abranches E;Silva M;Pradier L;Schulz H;Hummel O;Henrique D;Bekman E
从胚胎干细胞（ES）体外产生神经元是一种有前途的方法，以产生适合神经组织修复和神经系统的细胞替代疗法的细胞。促进ES细胞向神经谱系分化的现有方法试图以不同的方式复制胚胎神经发育的多步骤过程。然而，为了以有效和可重复的方式实现这一目标，需要更好地了解该过程中所涉及的细胞和分子事件，从神经上皮祖细胞的初始规格到其最终分化为神经元和神经胶质细胞。在这项工作中，我们的主要阶段和过渡时发生的ES细胞被驱动到一个神经的命运，使用贴壁单层培养系统的特点。我们建立了改进的条件，以常规地产生高度同质的神经上皮祖细胞培养物，这些祖细胞在获得神经元产生能力时组织成神经管样细胞。在roepithelium内，神经上皮祖细胞显示其胚胎对应物的形态和功能特征，即，顶端-基底极性、活跃的Notch信号传导以及神经元和神经胶质产生的适当时机。为了表征与神经发育的每个特定阶段相关的全局基因活性，通过微阵列分析确定在体外分化方案期间出现的不同细胞群体的全转录组。通过使用胚胎为导向的标准聚类的差异表达的基因，我们定义了五个基因表达的签名，相关的连续阶段的路径从ES细胞到神经元。这些包括ES细胞进入分化后出现的原始外胚层样阶段的基因签名，以及神经祖细胞发育的后续阶段的三个基因签名，从神经诱导后的早期阶段到终末分化前的最后阶段。总的来说，我们的工作证实并扩展了单层ES细胞神经分化和胚胎神经发育之间的细胞和分子相似性，揭示了从未定型细胞到分化神经元的多步骤过程的遗传网络的新方面。
The in vitro generation of neurons from embryonic stem (ES) cells is a promising approach to produce cells suitable for neural tissue repair and cell-based replacement therapies of the nervous system. Available methods to promote ES cell differentiation towards neural lineages attempt to replicate, in different ways, the multistep process of embryonic neural development. However, to achieve this aim in an efficient and reproducible way, a better knowledge of the cellular and molecular events that are involved in the process, from the initial specification of neuroepithelial progenitors to their terminal differentiation into neurons and glial cells, is required. In this work, we characterize the main stages and transitions that occur when ES cells are driven into a neural fate, using an adherent monolayer culture system. We established improved conditions to routinely produce highly homogeneous cultures of neuroepithelial progenitors, which organize into neural tube-like rosettes when they acquire competence for neuronal production. Within rosettes, neuroepithelial progenitors display morphological and functional characteristics of their embryonic counterparts, namely, apico-basal polarity, active Notch signalling, and proper timing of production of neurons and glia. In order to characterize the global gene activity correlated with each particular stage of neural development, the full transcriptome of different cell populations that arise during the in vitro differentiation protocol was determined by microarray analysis. By using embryo-oriented criteria to cluster the differentially expressed genes, we define five gene expression signatures that correlate with successive stages in the path from ES cells to neurons. These include a gene signature for a primitive ectoderm-like stage that appears after ES cells enter differentiation, and three gene signatures for subsequent stages of neural progenitor development, from an early stage that follows neural induction to a final stage preceding terminal differentiation. Overall, our work confirms and extends the cellular and molecular parallels between monolayer ES cell neural differentiation and embryonic neural development, revealing in addition novel aspects of the genetic network underlying the multistep process that leads from uncommitted cells to differentiated neurons.
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影响因子：
64.8
作者：
Brons, I. Gabrielle M.;Smithers, Lucy E.;Vallier, Ludovic
通讯作者：
Vallier, Ludovic
DOI：
10.1073/pnas.1734197100
发表时间：
2003-09-30
影响因子：
11.1
作者：
Aubert, J;Stavridis, MP;Smith, A
通讯作者：
Smith, A
影响因子：
9.8
作者：
通讯作者：
--
影响因子：
2.7
作者：
BAIN, G;KITCHENS, D;GOTTLIEB, DI
通讯作者：
GOTTLIEB, DI
DOI：
10.1111/j.2517-6161.1995.tb02031.x
发表时间：
1995-01-01
影响因子：
5.8
作者：
BENJAMINI, Y;HOCHBERG, Y
通讯作者：
HOCHBERG, Y