Trilck Michaela, Hübner Rayk, Frech Moritz J
Albrecht-Kossel-Institute for Neuroregeneration (AKos), University of Rostock, Gehlsheimer Straße 20, 18147, Rostock, Germany.
Methods Mol Biol. 2016;1353:233-59. doi: 10.1007/7651_2014_166.
Patient-specific induced pluripotent stem cells (iPSCs) are discussed to provide a powerful tool to investigate pathological mechanisms of diseases. Moreover, such cells might be a future platform for individualized personal treatment of diseases with a broad spectrum of mutations and thus resulting in phenotypical specificities.Here, we present a protocol for the induction of induced pluripotent stem cells from patient fibroblasts with Niemann-Pick type C1 disease (NPC1). The induction is based on a retroviral system, using the "classical" transcription factors, which were described by Takahashi and colleagues in 2007. To obtain a neuronal in vitro model system of NPC1, human iPSCs were differentiated to neural progenitor cells (NPCs) and subsequently to cells of the neural lineage, namely, neurons and glial cells. iPSCs, NPCs, and terminal neuronal differentiated cells (NDCs) were characterized by means of immunocytochemistry as well as patch clamp recordings and calcium imaging to prove the functional maturation.
患者特异性诱导多能干细胞（iPSCs）被认为是研究疾病病理机制的有力工具。此外，这类细胞可能成为未来个性化治疗多种具有广泛突变从而导致表型特异性疾病的平台。在此，我们展示了一种从患有尼曼-匹克C1型病（NPC1）的患者成纤维细胞诱导生成诱导多能干细胞的方案。诱导基于逆转录病毒系统，使用高桥等人在2007年描述的“经典”转录因子。为获得NPC1的神经元体外模型系统，将人iPSCs分化为神经祖细胞（NPCs），随后再分化为神经谱系细胞，即神经元和神经胶质细胞。通过免疫细胞化学、膜片钳记录和钙成像对iPSCs、NPCs和终末神经元分化细胞（NDCs）进行表征，以证明其功能成熟。
