Maturation of Human Pluripotent Stem Cell-Derived Cerebellar Neurons in the Absence of Co-culture
Maturation of Human Pluripotent Stem Cell-Derived Cerebellar Neurons in the Absence of Co-culture
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DOI：
10.3389/fbioe.2020.00070
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发表时间：
2020-02-14
影响因子：
5.7
通讯作者：
Carmo-Fonseca, Maria
中科院分区：
文献类型：
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作者：
Silva, Teresa P.;Bekman, Evguenia P.;Carmo-Fonseca, Maria
小脑在所有脊椎动物中起着至关重要的作用，许多神经系统疾病都与小脑功能障碍有关。小脑研究的一个主要限制是缺乏足够的疾病模型。作为动物模型的替代方案，已使用从多能干细胞分化的小脑神经元。然而，以前的研究只产生了有限数量的浦肯野细胞。此外，浦肯野细胞的体外产生需要共培养系统，这可能会将未知成分引入系统。在这里，我们描述了一种新的分化策略，使用确定的培养基产生浦肯野细胞，颗粒细胞，中间神经元，和小脑深核投射神经元，自我形成和分化成电活性细胞。使用一个确定的基础培养基优化神经元细胞培养，我们成功地促进小脑前体细胞的分化，而不需要共培养。我们预计，我们的研究结果可能有助于开发更好的模型，用于研究小脑功能障碍，同时为治疗小脑退行性疾病的自体替代策略的发展提供了一个进步。
The cerebellum plays a critical role in all vertebrates, and many neurological disorders are associated with cerebellum dysfunction. A major limitation in cerebellar research has been the lack of adequate disease models. As an alternative to animal models, cerebellar neurons differentiated from pluripotent stem cells have been used. However, previous studies only produced limited amounts of Purkinje cells. Moreover, in vitro generation of Purkinje cells required co-culture systems, which may introduce unknown components to the system. Here we describe a novel differentiation strategy that uses defined medium to generate Purkinje cells, granule cells, interneurons, and deep cerebellar nuclei projection neurons, that self-formed and differentiated into electrically active cells. Using a defined basal medium optimized for neuronal cell culture, we successfully promoted the differentiation of cerebellar precursors without the need for co-culturing. We anticipate that our findings may help developing better models for the study of cerebellar dysfunctions, while providing an advance toward the development of autologous replacement strategies for treating cerebellar degenerative diseases.