Silva Teresa P, Bekman Evguenia P, Fernandes Tiago G, Vaz Sandra H, Rodrigues Carlos A V, Diogo Maria Margarida, Cabral Joaquim M S, Carmo-Fonseca Maria
Instituto de Medicina Molecular João Lobo Antunes, Faculdade de Medicina, Universidade de Lisboa, Lisbon, Portugal.
Department of Bioengineering and Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal.
Front Bioeng Biotechnol. 2020 Feb 14;8:70. doi: 10.3389/fbioe.2020.00070. eCollection 2020.
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, 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.
小脑在所有脊椎动物中都起着关键作用，许多神经疾病都与小脑功能障碍有关。小脑研究的一个主要局限在于缺乏合适的疾病模型。作为动物模型的替代方法，已经使用了从多能干细胞分化而来的小脑神经元。然而，先前的研究仅产生了有限数量的浦肯野细胞。此外，浦肯野细胞的生成需要共培养系统，这可能会将未知成分引入该系统。在这里，我们描述了一种新的分化策略，该策略使用特定培养基生成浦肯野细胞、颗粒细胞、中间神经元和小脑深部核团投射神经元，这些细胞能自我形成并分化为电活动细胞。使用针对神经元细胞培养优化的特定基础培养基，我们成功促进了小脑前体细胞的分化，而无需共培养。我们预计，我们的发现可能有助于开发更好的模型来研究小脑功能障碍，同时为治疗小脑退行性疾病的自体替代策略的发展提供进展。
