Mechanobiology Institute Singapore, National University of Singapore, T-Lab, #05-01, 5A Engineering Drive 1, Singapore, 117411, Singapore.
Institute of Medical Biology, 8A Biomedical Grove, #06-06 Immunos, Singapore, 138648, Singapore.
Sci Rep. 2018 Jun 22;8(1):9567. doi: 10.1038/s41598-018-27653-1.
Parkinson's Disease is a progressive neurodegenerative disorder attributed to death of mesencephalic dopaminergic (DA) neurons. Pluripotent stem cells have great potential in the study for this late-onset disease, but acquirement of cells that are robust in quantity and quality is still technically demanding. Biophysical cues have been shown to direct stem cell fate, but the effect of different topographies in the lineage commitment and subsequent maturation stages of cells have been less examined. Using human induced pluripotent stem cells (iPSCs), we applied topographical patterns sequentially during differentiation stages and examined their ability to influence derivation yield and functionality of regionalized subtype-specific DA neurons. Gratings showed higher yield of DA neurons and may be beneficial for initial lineage commitment. Cells derived on pillars in the terminal differentiation stage have increased neuronal complexity, and were more capable of firing repetitive action potentials, showing that pillars yielded better network formation and functionality. Our topography platform can be applied to patient-derived iPSCs as well, and that cells harbouring LRRK2 mutation were more functionally mature when optimal topographies were applied sequentially. This will hopefully accelerate development of robust cell models that will provide novel insights into discovering new therapeutic approaches for Parkinson's Disease.
帕金森病是一种进行性神经退行性疾病，归因于中脑多巴胺能（DA）神经元的死亡。多能干细胞在这种迟发性疾病的研究中有很大的潜力，但获得大量且质量优良的细胞在技术上仍然具有挑战性。生物物理线索已被证明可以指导干细胞命运，但不同拓扑结构在细胞谱系决定和随后的成熟阶段的影响尚未得到充分研究。使用人诱导多能干细胞（iPSCs），我们在分化阶段顺序施加拓扑图案，并检查它们影响区域性亚型特异性 DA 神经元的衍生产量和功能的能力。光栅显示出更高的 DA 神经元产量，可能有利于初始谱系决定。在终末分化阶段在柱上衍生的细胞具有增加的神经元复杂性，并且更能够发射重复动作电位，表明柱产生了更好的网络形成和功能。我们的拓扑平台也可以应用于患者来源的 iPSCs，并且当最佳拓扑结构顺序施加时，携带 LRRK2 突变的细胞具有更高的功能成熟度。这有望加速开发稳健的细胞模型，为发现治疗帕金森病的新方法提供新的见解。
