Creation of a library of induced pluripotent stem cells from Parkinsonian patients

Abstract

Induced pluripotent stem cells (iPSCs) are becoming an important source of pre-clinical models for research focusing on neurodegeneration. They offer the possibility for better understanding of common and divergent pathogenic mechanisms of brain diseases. Moreover, iPSCs provide a unique opportunity to develop personalized therapeutic strategies, as well as explore early pathogenic mechanisms, since they rely on the use of patients’ own cells that are otherwise accessible only post-mortem, when neuronal death-related cellular pathways and processes are advanced and adaptive. Neurodegenerative diseases are in majority of unknown cause, but mutations in specific genes can lead to familial forms of these diseases. For example, mutations in the superoxide dismutase 1 gene lead to the motor neuron disease amyotrophic lateral sclerosis (ALS), while mutations in the SNCA gene encoding for alpha-synuclein protein lead to familial Parkinson’s disease (PD). The generations of libraries of familial human ALS iPSC lines have been described, and the iPSCs rapidly became useful models for studying cell autonomous and non-cell autonomous mechanisms of the disease. Here we report the generation of a comprehensive library of iPSC lines of familial PD and an associated synucleinopathy, multiple system atrophy (MSA). In addition, we provide examples of relevant neural cell types these iPSC can be differentiated into, and which could be used to further explore early disease mechanisms. These human cellular models will be a valuable resource for identifying common and divergent mechanisms leading to neurodegeneration in PD and MSA.

PSEN1 Mutant iPSC-Derived Model Reveals Severe Astrocyte Pathology in Alzheimer's Disease

“…Fibroblasts and peripheral blood mononuclear cells T cells were isolated and cultured as previously described ( Korhonen et al., 2015 , Qu et al., 2013 ). Somatic cells were reprogrammed to iPSCs with either lentiviral vectors, CytoTune -iPS 1.0, or CytoTune -iPS 2.0 Sendai Reprogramming Kits (Invitrogen) as previously described ( Holmqvist et al., 2016 ). iPSCs were grown on Matrigel-coated (Corning) plates in Essential 8 Medium (E8; Life Technologies) and passaged with 0.5 mM EDTA in the presence of 5 μM Y-27632 ROCK inhibitor (Selleckchem).…”

Structural Immaturity of Human iPSC-Derived Cardiomyocytes: In Silico Investigation of Effects on Function and Disease Modeling

“…Written informed consent was obtained from the donor. Skin biopsy derived fibroblasts were reprogrammed with CytoTune® iPS Sendai Reprogramming kit (Thermo Scientific, MA, USA) as previously described (Holmqvist et al, 2016 ), with slight modifications. Briefly, fibroblasts (1 × 10 5 ) were transduced with 3 or 4 separate vectors including the four Yamanaka factors OCT-3/4, KLF-4, SOX-2 and c-MYC .…”

Astrocyte alterations in neurodegenerative pathologies and their modeling in human induced pluripotent stem cell platforms

“…The majority of the current human data from neurodegenerative diseases originates from post-mortem samples, representing thus only the end stage of the disease. Novel methodologies have enabled the generation of comprehensive iPSC cell libraries [ 207 ], which will in the future be an invaluable model resource for studying neurodegeneration. For example, iPSC-based studies have shown aberrant morphological changes in AD astrocytes [ 208 ], as well as APOE-related neurotrophic and synaptic disturbances [ 19 ].…”