Autor: Christopher W. Vaughan, Ernst J. Wolvetang, Carolyn M. Sue, Nicholas F. Blair, Paula Woodbridge, Stephen Gao
Rok vydání: 2014
Předmět:
Zdroj: Journal of Clinical Neuroscience. 21:2056
ISSN: 0967-5868
DOI: 10.1016/j.jocn.2014.06.088
Popis: Parkinson’s disease (PD) is a common neurodegenerative disorder with no current treatment that arrests the progress of the disease. The objective of this study was to develop cellular models of PD for use in the study of the pathophysiological pathways involved in the disease. We aimed to generate these by deriving induced-pluripotent stem (iPS) cells from patients with monogenic forms of the disease and then differentiating these stem cells into dopaminergic neurons that would each carry the causative mutations. We generated fibroblast cultures from skin biopsies taken from five patients with monogenic forms of PD. iPS cell lines were produced using lentiviral transduction of the four Yamanaka factors. The iPS cells were differentiated into dopaminergic neurons using established protocols. iPS cell lines were generated from all five patients. They were confirmed to be pluripotent using standard assays. Each iPS cell line was successfully differentiated into midbrain dopaminergic neurons as confirmed by the presence of markers Lmx1 and FoxA2 on immunocytochemistry. The relative yield of neurons generated by each cell line was studied using Western blotting and this confirmed a comparable efficiency of disease and control cell lines. Neurophysiological studies were performed using whole cell patch-clamping and this confirmed the presence of action potentials with characteristic features of dopaminergic neurons. We demonstrate that the neuronal differentiation of iPS cells is a feasible approach for generating patient-derived tissue-specific cell models of PD.
Databáze: OpenAIRE
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