| Autor: |
Stefanie, Traub, Ralf, Heilker |
| Rok vydání: |
2019 |
| Předmět: |
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| Zdroj: |
Methods in molecular biology (Clifton, N.J.). 1994 |
| ISSN: |
1940-6029 |
| Popis: |
Human induced pluripotent stem (hiPS) cell-derived neurons promise to provide better model cells for drug discovery in the context of neurodegenerative and neuropsychiatric diseases. The neuronal differentiation protocol described encompasses a cellular amplification phase for hiPS-derived neural progenitor (NP) cells. Thus, the combination of growth factor-driven expansion and inhibition of notch (GRINCH) enabled the scalable production of neurons in sufficient numbers to meet the immense material needs of a high-throughput screening (HTS) campaign. These GRINCH cells matured in 384-well microplates display neuronal markers and electrophysiological activity. The differentiation protocol was applicable to various human hiPS cell clones. In a finding and profiling campaign for modulators of the tropomyosin receptor kinase B (TrkB), the GRINCH neurons were shown to be suitable for measuring the phosphorylation and downstream signaling of the endogenously expressed TrkB. The employed techniques in the amplified luminescent proximity homogeneous assay (Alpha) and the high-throughput reverse transcription polymerase chain reaction (RT-PCR) format are transferable to other pharmaceutical drug targets. Together with the GRINCH neurons, these detection technologies open new experimental routes with tremendous potential for early drug discovery. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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