Neurogenomics Division, The Translational Genomics Research Institute (TGen), Phoenix, AZ, USA.
Neurosci Lett. 2011 Sep 20;502(3):219-24. doi: 10.1016/j.neulet.2011.07.048. Epub 2011 Aug 4.
Human induced pluripotent stem cells (iPSCs) have become an intriguing approach for neurological disease modeling, because neural lineage-specific cell types that retain the donors' complex genetics can be established in vitro. The statistical power of these iPSC-based models, however, is dependent on accurate diagnoses of the somatic cell donors; unfortunately, many neurodegenerative diseases are commonly misdiagnosed in live human subjects. Postmortem histopathological examination of a donor's brain, combined with premortem clinical criteria, is often the most robust approach to correctly classify an individual as a disease-specific case or unaffected control. In this study, we describe iPSCs generated from a skin biopsy collected postmortem during the rapid autopsy of a 75-year-old male, whole body donor, defined as an unaffected neurological control by both clinical and histopathological criteria. These iPSCs were established in a feeder-free system by lentiviral transduction of the Yamanaka factors, Oct3/4, Sox2, Klf4, and c-Myc. Selected iPSC clones expressed both nuclear and surface antigens recognized as pluripotency markers of human embryonic stem cells (hESCs) and were able to differentiate in vitro into neurons and glia. Statistical analysis also demonstrated that fibroblast proliferation was significantly affected by biopsy site, but not donor age (within an elderly cohort). These results provide evidence that autopsy donor-derived fibroblasts can be successfully reprogrammed into iPSCs, and may provide an advantageous approach for generating iPSC-based neurological disease models.
人类诱导多能干细胞(iPSC)已成为神经疾病建模的一种有趣方法,因为可以在体外建立保留供体复杂遗传信息的神经谱系特异性细胞类型。然而,这些基于 iPSC 的模型的统计能力取决于对体细胞供体的准确诊断;不幸的是,许多神经退行性疾病在活体人类受试者中经常被误诊。对供体大脑进行死后组织病理学检查,并结合生前临床标准,通常是正确分类个体为特定疾病病例或未受影响对照的最可靠方法。在这项研究中,我们描述了从一位 75 岁男性的死后快速尸检中收集的皮肤活检中生成的 iPSC,该男性被临床和组织病理学标准定义为未受影响的神经对照。这些 iPSC 是通过慢病毒转导 Yamanaka 因子(Oct3/4、Sox2、Klf4 和 c-Myc)在无饲养细胞的系统中建立的。选定的 iPSC 克隆表达了核和表面抗原,这些抗原被认为是人类胚胎干细胞(hESC)的多能性标记物,并且能够在体外分化为神经元和神经胶质细胞。统计分析还表明,成纤维细胞的增殖受到活检部位的显著影响,但不受供体年龄(在老年队列中)的影响。这些结果提供了证据,证明尸检供体来源的成纤维细胞可以成功地被重编程为 iPSC,并且可能为生成基于 iPSC 的神经疾病模型提供了有利的方法。