ASCL1- and DLX2-induced GABAergic neurons from hiPSC-derived NPCs
| Autor: | Samuel K. Powell, Erin Flaherty, Michael B. Fernando, Jubao Duan, Hanwen Zhang, Paul A. Slesinger, Seok-Man Ho, Siwei Zhang, Natalie Barretto, Kristen J. Brennand |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
education.field_of_study Somatic cell General Neuroscience Population Context (language use) Biology Neural stem cell Article 03 medical and health sciences ASCL1 030104 developmental biology 0302 clinical medicine Directed differentiation GABAergic education Reprogramming Neuroscience 030217 neurology & neurosurgery |
| Zdroj: | J Neurosci Methods |
| ISSN: | 1872-678X |
| Popis: | Background Somatic cell reprogramming is routinely used to generate donor-specific human induced pluripotent stem cells (hiPSCs) to facilitate studies of disease in a human context. The directed differentiation of hiPSCs can generate large quantities of patient-derived cells; however, such methodologies frequently yield heterogeneous populations of neurons and glia that require extended timelines to achieve electrophysiological maturity. More recently, transcription factor-based induction protocols have been show to rapidly generate defined neuronal populations from hiPSCs. New method In a manner similar to our previous adaption of NGN2-glutamatergic neuronal induction from hiPSC-derived neural progenitor cells (NPCs), we now adapt an established protocol of lentiviral overexpression of ASCL1 and DLX2 to hiPSC-NPCs. Results We demonstrate induction of a robust and highly pure population of functional GABAergic neurons (iGANs). Importantly, we successfully applied this technique to hiPSC-NPCs derived from ten donors across two independent laboratories, finding it to be an efficient and highly reproducible approach to generate induced GABAergic neurons. Our results show that, like hiPSC-iGANs, NPC-iGANs exhibit increased GABAergic marker expression, electrophysiological maturity, and have distinct transcriptional profiles that distinguish them from other cell-types of the brain. Nonetheless, until donor-matched hiPSCs-iGANs and NPC-iGANs are directly compared, we cannot rule out the possibility that subtle differences in patterning or maturity may exist between these populations; one should always control for cell source in all iGAN experiments. Conclusions This methodology, relying upon an easily cultured starting population of hiPSC-NPCs, makes possible the generation of large-scale defined co-cultures of induced glutamatergic and GABAergic neurons for hiPSC-based disease models and precision drug screening. |
| Databáze: | OpenAIRE |
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