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.

中文翻译：

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鉴定人神经干细胞的胶质生成状态以优化体外星形胶质细胞分化

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用于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 hNSC 对人类胶质发生进行了转录表征。基于这些发现，我们建立了一种快速高效的成熟人星形胶质细胞分化方案。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

我们发现 NSC 越来越倾向于产生具有重复细胞传代的星形胶质细胞。NSCs 的胶质生成表型以干细胞因子（例如 SOX1、SOX2、EGFR）的下调和神经胶质相关因子（例如 NFIX、SOX9、PDGFRa）的增加为标志。使用晚期通道神经干细胞，可以在 28 天内实现快速和强大的星形胶质细胞分化。

与现有方法的比较

在已发表的协议中，成熟的星形胶质细胞通常需要大约三个月的时间才能产生。在体外生成星形胶质细胞的难度、费用和时间是神经胶质细胞研究的主要障碍。我们表明可以在 28 天内实现快速和强大的星形胶质细胞分化。我们在此通过对 hNSC 进行广泛的顺序转录组分析来描述新型胶质细胞生成干细胞群特征的特征。转录组特征可能有助于识别适当的分裂成熟度。

结论

这项工作阐明了与 NSC 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生成并开发有助于研究中枢神经系统疾病（如多发性硬化症）的新型临床前模型。