Parkinson's Disease Patient-Derived Induced Pluripotent Stem Cells Free of Viral Reprogramming Factors

Induced pluripotent stem cells (iPSCs) derived from somatic cells of patients represent a powerful tool for biomedical research and may provide a source for replacement therapies. However, the use of viruses encoding the reprogramming factors represents a major limitation of the current technology since even low vector expression may alter the differentiation potential of the iPSCs or induce malignant transformation. Here, we show that fibroblasts from five patients with idiopathic Parkinson's disease can be efficiently reprogrammed and subsequently differentiated into dopaminergic neurons. Moreover, we derived hiPSCs free of reprogramming factors using Cre-recombinase excisable viruses. Factor-free hiPSCs maintain a pluripotent state and show a global gene expression profile, more closely related to hESCs than to hiPSCs carrying the transgenes. Our results indicate that residual transgene expression in virus-carrying hiPSCs can affect their molecular characteristics and that factor-free hiPSCs therefore represent a more suitable source of cells for modeling of human disease.
Howard Hughes Medical Institute (Collaborative Innovation Award)
Life Sciences Research Foundation (Merck Fellow)
Michael Stern Parkinson's Research Foundation
Morris K. Udall Center for Excellence in Parkinson’s Research (grant P50NS39793)
National Institutes of Health (U.S.) (NIH grant R37-CA084198)
National Institutes of Health (U.S.) (NIH grant RO1-CA087869)
National Institutes of Health (U.S.) (grant NIH RO1-HD045022)