Homogenous generation of dopaminergic neurons from multiple hiPSC lines by transient expression of transcription factors
| Autor: | Anupam Raina, Mathias Bähr, Sebastian Kügler, Sameehan Mahajani, Claudia Fokken |
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| Rok vydání: | 2019 |
| Předmět: |
Adult
Cancer Research Tyrosine 3-Monooxygenase Dopamine Parkinson's disease Genetic Vectors Green Fluorescent Proteins Induced Pluripotent Stem Cells Immunology Stem-cell differentiation Context (language use) Substantia nigra Biology hiPSC Article Cell Line 03 medical and health sciences Cellular and Molecular Neuroscience 0302 clinical medicine medicine Animals Humans lcsh:QH573-671 Induced pluripotent stem cell Transcription factor 030304 developmental biology 0303 health sciences Reporter gene lcsh:Cytology Pars compacta Dopaminergic Neurons Neurodegeneration Dopaminergic Cell Differentiation Cell Biology Dependovirus medicine.disease Rats 3. Good health nervous system alpha-Synuclein Female Neuroscience 030217 neurology & neurosurgery Transcription Factors |
| Zdroj: | Cell Death & Disease Cell Death and Disease, Vol 10, Iss 12, Pp 1-15 (2019) |
| ISSN: | 2041-4889 |
| DOI: | 10.1038/s41419-019-2133-9 |
| Popis: | A major hallmark of Parkinson's disease is loss of dopaminergic neurons in the substantia nigra pars compacta (SNpc). The pathophysiological mechanisms causing this relatively selective neurodegeneration are poorly understood, and thus experimental systems allowing to study dopaminergic neuron dysfunction are needed. Induced pluripotent stem cells (iPSCs) differentiated toward a dopaminergic neuronal phenotype offer a valuable source to generate human dopaminergic neurons. However, currently available protocols result in a highly variable yield of dopaminergic neurons depending on the source of hiPSCs. We have now developed a protocol based on HBA promoter-driven transient expression of transcription factors by means of adeno-associated viral (AAV) vectors, that allowed to generate very consistent numbers of dopaminergic neurons from four different human iPSC lines. We also demonstrate that AAV vectors expressing reporter genes from a neuron-specific hSyn1 promoter can serve as surrogate markers for maturation of hiPSC-derived dopaminergic neurons. Dopaminergic neurons differentiated by transcription factor expression showed aggravated neurodegeneration through α-synuclein overexpression, but were not sensitive to γ-synuclein overexpression, suggesting that these neurons are well suited to study neurodegeneration in the context of Parkinson’s disease. |
| Databáze: | OpenAIRE |
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