Abstract

The generation of disease-specific induced pluripotent stem cell (iPSC) lines from patients with incurable diseases is a promising approach for studying disease mechanisms and drug screening. Such innovation enables to obtain autologous cell sources in regenerative medicine. Herein, we report the generation and characterization of iPSCs from fibroblasts of patients with sporadic or familial diseases, including Parkinson's disease (PD), Alzheimer's disease (AD), juvenile-onset, type I diabetes mellitus (JDM), and Duchenne type muscular dystrophy (DMD), as well as from normal human fibroblasts (WT). As an example to modeling disease using disease-specific iPSCs, we also discuss the previously established childhood cerebral adrenoleukodystrophy (CCALD)- and adrenomyeloneuropathy (AMN)-iPSCs by our group. Through DNA fingerprinting analysis, the origins of generated disease-specific iPSC lines were identified. Each iPSC line exhibited an intense alkaline phosphatase activity, expression of pluripotent markers, and the potential to differentiate into all three embryonic germ layers: the ectoderm, endoderm, and mesoderm. Expression of endogenous pluripotent markers and downregulation of retrovirus-delivered transgenes [OCT4 (POU5F1), SOX2, KLF4, and c-MYC] were observed in the generated iPSCs. Collectively, our results demonstrated that disease-specific iPSC lines characteristically resembled hESC lines. Furthermore, we were able to differentiate PD-iPSCs, one of the disease-specific-iPSC lines we generated, into dopaminergic (DA) neurons, the cell type mostly affected by PD. These PD-specific DA neurons along with other examples of cell models derived from disease-specific iPSCs would provide a powerful platform for examining the pathophysiology of relevant diseases at the cellular and molecular levels and for developing new drugs and therapeutic regimens.

Highlights

  • Human embryonic stem cells are isolated from the inner cell mass of the developing blastocyst, indefinitely maintaining their capability for self-renewal and pluripotent nature (Thomson et al, 1998)

  • As a breakthrough discovery in regenerative medicine, adult human fibroblasts have been successfully reprogrammed via overexpression of pluripotency-related transcription factors, which include octamer-binding protein 4 (OCT4) (POU5F1), SOX2, Kruppel-like factor 4 (KLF4), and c-myelocytomatosis oncogene (MYC), to establish human induced pluripotent stem cells (Takahashi et al, 2007)

  • Patient-derived disease-specific fibroblasts and normal neonatal human dermal fibroblasts were reprogrammed by retroviral-mediated delivery of four defined factors (OCT4, SOX2, KLF4, and c-MYC) (Table 1)

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Summary

Introduction

Human embryonic stem cells (hESCs) are isolated from the inner cell mass of the developing blastocyst, indefinitely maintaining their capability for self-renewal and pluripotent nature (Thomson et al, 1998). The disease-specific hiPSCs can be differentiated into cell-types of their relevant diseases. We generated PD-specific DA neurons from the hiPSCs derived from fibroblasts of PD patients.

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