Identification of the gliogenic state of human neural stem cells to optimize in vitro astrocyte differentiation
Identification of the gliogenic state of human neural stem cells to optimize in vitro astrocyte differentiation
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DOI：
10.1016/j.jneumeth.2021.109284
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发表时间：
2021-07-21
影响因子：
3
通讯作者：
Siffrin, Volker
中科院分区：
文献类型：
--
作者：
Alisch, Marlen;Kerkering, Janis;Siffrin, Volker
背景：人类临床前模型对于推进生物医学研究至关重要。特别是在人类系统中星形胶质细胞分化的一致和强大的协议是罕见的。新方法：我们使用胚胎H9衍生的hNSCs进行了人类胶质细胞生成的转录表征。基于这些发现，我们建立了一个快速，高效的方案，成熟的人星形胶质细胞的分化。我们可以在诱导多能干细胞（iPSC）衍生的NSC中重现这些结果。结果：我们确定了一个增加的倾向，神经干细胞产生星形胶质细胞与反复细胞传代。神经干细胞的胶质细胞生成表型以干细胞因子（例如SOX 1、SOX 2、EGFR）的下调和胶质细胞相关因子（例如NFIX、SOX 9、PDGFRa）的增加为标志。使用晚期传代的NSC，可以在28天内实现快速和稳健的星形胶质细胞分化。与现有方法的比较：在已发表的方案中，通常需要大约三个月才能产生成熟的星形胶质细胞。体外培养星形胶质细胞的困难、费用和时间是胶质细胞研究的主要障碍。我们表明，快速和强大的星形胶质细胞分化可以在28天内实现。在这里，我们描述了一个广泛的连续的转录组分析的hNSCs的特征的一种新的神经胶质干细胞群的签名。转录组签名可能有助于识别适当的部门成熟度。结论：这项工作揭示了与神经干细胞快速分化为神经胶质细胞相关的因素。这些发现有助于了解人类胶质细胞生成和开发新的临床前模型，这将有助于研究中枢神经系统疾病，如多发性硬化症。
Background: Human preclinical models are crucial for advancing biomedical research. In particular consistent and robust protocols for astrocyte differentiation in the human system are rare. New method: We performed a transcriptional characterization of human gliogenesis using embryonic H9- derived hNSCs. Based on these findings we established a fast and highly efficient protocol for the differentiation of mature human astrocytes. We could reproduce these results in induced pluripotent stem cell (iPSC)-derived NSCs. Results: We identified an increasing propensity of NSCs to give rise to astrocytes with repeated cell passaging. The gliogenic phenotype of NSCs was marked by a down-regulation of stem cell factors (e.g. SOX1, SOX2, EGFR) and an increase of glia-associated factors (e.g. NFIX, SOX9, PDGFRa). Using late passage NSCs, rapid and robust astrocyte differentiation can be achieved within 28 days. Comparison with existing method(s): In published protocols it usually takes around three months to yield in mature astrocytes. The difficulty, expense and time associated with generating astrocytes in vitro represents a major roadblock for glial cell research. We show that rapid and robust astrocyte differentiation can be achieved within 28 days. We describe here by an extensive sequential transcriptome analysis of hNSCs the characterization of the signature of a novel gliogenic stem cell population. The transcriptomic signature might serve to identify the proper divisional maturity. Conclusions: This work sheds light on the factors associated with rapid NSC differentiation into glial cells. These findings contribute to understand human gliogenesis and to develop novel preclinical models that will help to study CNS disease such as Multiple Sclerosis.