Magistri Marco, Khoury Nathalie, Mazza Emilia Maria Cristina, Velmeshev Dmitry, Lee Jae K, Bicciato Silvio, Tsoulfas Pantelis, Faghihi Mohammad Ali
Department of Psychiatry and Behavioral Sciences, Center for Therapeutic Innovation, University of Miami Miller School of Medicine, 1501 NW 10th Ave, BRB 508, Miami, FL, 33136, USA.
Department of Life Sciences, Center for Genome Research, University of Modena and Reggio Emilia, Modena, Italy.
Eur J Neurosci. 2016 Nov;44(10):2858-2870. doi: 10.1111/ejn.13382. Epub 2016 Sep 25.
Astrocytes are a morphologically and functionally heterogeneous population of cells that play critical roles in neurodevelopment and in the regulation of central nervous system homeostasis. Studies of human astrocytes have been hampered by the lack of specific molecular markers and by the difficulties associated with purifying and culturing astrocytes from adult human brains. Human neural progenitor cells (NPCs) with self-renewal and multipotent properties represent an appealing model system to gain insight into the developmental genetics and function of human astrocytes, but a comprehensive molecular characterization that confirms the validity of this cellular system is still missing. Here we used an unbiased transcriptomic analysis to characterize in vitro culture of human NPCs and to define the gene expression programs activated during the differentiation of these cells into astrocytes using FBS or the combination of CNTF and BMP4. Our results demonstrate that in vitro cultures of human NPCs isolated during the gliogenic phase of neurodevelopment mainly consist of radial glial cells (RGCs) and glia-restricted progenitor cells. In these cells the combination of CNTF and BMP4 activates the JAK/STAT and SMAD signaling cascades, leading to the inhibition of oligodendrocytes lineage commitment and activation of astrocytes differentiation. On the other hand, FBS-derived astrocytes have properties of reactive astrocytes. Our work suggests that in vitro culture of human NPCs represents a valuable cellular system to study human disorders characterized by impairment of astrocytes development and function. Our datasets represent an important resource for researchers studying human astrocytes development and might set the basis for the discovery of novel human-specific astrocyte markers.
星形胶质细胞是一群形态和功能各异的细胞,在神经发育和中枢神经系统稳态调节中发挥着关键作用。由于缺乏特异性分子标记以及从成人人类大脑中纯化和培养星形胶质细胞存在困难,对人类星形胶质细胞的研究受到了阻碍。具有自我更新和多能特性的人类神经祖细胞(NPCs)是一个有吸引力的模型系统,有助于深入了解人类星形胶质细胞的发育遗传学和功能,但仍缺乏全面的分子表征来证实这个细胞系统的有效性。在这里,我们使用无偏转录组分析来表征人类NPCs的体外培养,并使用胎牛血清(FBS)或睫状神经营养因子(CNTF)与骨形态发生蛋白4(BMP4)的组合来定义这些细胞分化为星形胶质细胞过程中激活的基因表达程序。我们的结果表明,在神经发育的胶质生成阶段分离的人类NPCs的体外培养主要由放射状胶质细胞(RGCs)和胶质限制祖细胞组成。在这些细胞中,CNTF和BMP4的组合激活了JAK/STAT和SMAD信号级联,导致少突胶质细胞谱系定向的抑制和星形胶质细胞分化的激活。另一方面,FBS来源的星形胶质细胞具有反应性星形胶质细胞的特性。我们的工作表明,人类NPCs的体外培养是一个有价值的细胞系统,可用于研究以星形胶质细胞发育和功能受损为特征的人类疾病。我们的数据集是研究人类星形胶质细胞发育的研究人员的重要资源,并可能为发现新的人类特异性星形胶质细胞标记奠定基础。