Inhibition of DNA methyltransferases and histone deacetylases induces astrocytic differentiation of neural progenitors.
Inhibition of DNA methyltransferases and histone deacetylases induces astrocytic differentiation of neural progenitors.
复制标题
DOI：
10.1016/j.scr.2013.03.003
复制
发表时间：
2013-07
影响因子：
1.2
通讯作者：
A. Majumder;S. Dhara;R. Swetenburg;M. Mithani;Kaixiang Cao;M. Medrzycki;Yuhong Fan;S. Stice
中科院分区：
文献类型：
--
作者：
A. Majumder;S. Dhara;R. Swetenburg;M. Mithani;Kaixiang Cao;M. Medrzycki;Yuhong Fan;S. Stice
了解如何指定人神经祖细胞向富集的非转化的人星形胶质细胞祖细胞的快速分化将提供一个关键的细胞来源，以进一步了解星形胶质细胞如何在神经功能和发育中发挥关键作用。来源于多能胚胎干细胞并在贴壁无血清培养物中繁殖的人神经祖细胞为神经细胞的快速产生提供了命运受限的可再生来源;然而，此类细胞高度折射星形细胞发生并显示出强烈的神经发生性偏好，类似于来自早期胚胎中枢神经系统（CNS）的神经祖细胞。我们发现，几个星形胶质细胞基因在这些祖细胞中高甲基化，可能阻止星形胶质细胞的产生，并导致这些祖细胞的前神经元命运。然而，通过氮杂胞苷（Aza-C）和曲古抑菌素A（TSA）的表观遗传修饰，以及神经祖细胞培养物中来自BMP 2和LIF的伴随信号传导改变了这种偏倚，导致早在分化的5天星形胶质细胞标记物的表达，神经元分化几乎完全抑制。所得细胞表达主要的星形胶质细胞标志物，适合与神经元共培养，可以作为星形胶质细胞祖细胞繁殖，并且是可冷冻保存的。虽然先前的报道已经从多能细胞产生星形胶质细胞，但分化需要基于细胞表面抗原的广泛培养或选择。无标记和快速分化过程的开发将加速未来从各种来源的多能细胞中衍生星形胶质细胞，包括但不限于与各种神经系统疾病相关的人类星形胶质细胞。
Understanding how to specify rapid differentiation of human neural progenitor towards enriched non-transformed human astrocyte progenitors will provide a critical cell source to further our understanding of how astrocytes play a pivotal role in neural function and development. Human neural progenitors derived from pluripotent embryonic stem cells and propagated in adherent serum-free cultures provide a fate restricted renewable source for quick production of neural cells; however, such cells are highly refractive to astrocytogenesis and show a strong neurogenic bias, similar to neural progenitors from the early embryonic central nervous system (CNS). We found that several astrocytic genes are hypermethylated in such progenitors potentially preventing generation of astrocytes and leading to the proneuronal fate of these progenitors. However, epigenetic modification by Azacytidine (Aza-C) and Trichostatin A (TSA), with concomitant signaling from BMP2 and LIF in neural progenitor cultures shifts this bias, leading to expression of astrocytic markers as early as 5days of differentiation, with near complete suppression of neuronal differentiation. The resultant cells express major astrocytic markers, are amenable to co-culture with neurons, can be propagated as astrocyte progenitors and are cryopreservable. Although previous reports have generated astrocytes from pluripotent cells, the differentiation required extensive culture or selection based on cell surface antigens. The development of a label free and rapid differentiation process will expedite future derivation of astrocytes from various sources pluripotent cells including, but not limited to, human astrocytes associated with various neurological diseases.