Nongenetic methodologies to reduce undesirable proliferation would be valuable when generating dopamine neurons from stem cells for transplantation in Parkinson's disease (PD). To this end, we modified an established method for controlled differentiation of human induced pluripotent stem cells (iPSCs) into midbrain dopamine neurons using two distinct methods: omission of FGF8 or the in‐process use of the DNA cross‐linker mitomycin‐C (MMC). We transplanted the cells to athymic rats with unilateral 6‐hydroxydopamine lesions and monitored long‐term survival and function of the grafts. Transplants of cells manufactured using MMC had low proliferation while still permitting robust survival and function comparable to that seen with transplanted dopamine neurons derived using genetic drug selection. Conversely, cells manufactured without FGF8 survived transplantation but exhibited poor in vivo function. Our results suggest that MMC can be used to reduce the number of proliferative cells in stem cell‐derived postmitotic neuron preparations for use in PD cell therapy.

中文翻译：

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诱导多能干细胞衍生的多巴胺神经元分化过程中的丝裂霉素-C治疗可减少增殖而不影响体内存活或功能

当从干细胞生成多巴胺神经元用于帕金森病 (PD) 的移植时，减少不良增殖的非遗传方法将是有价值的。为此，我们使用两种不同的方法修改了一种既定的方法，用于将人诱导多能干细胞 (iPSC) 控制分化为中脑多巴胺神经元：省略 FGF8 或在过程中使用 DNA 交联剂丝裂霉素-C (MMC) ）。我们将这些细胞移植到有单侧 6-羟基多巴胺损伤的无胸腺大鼠身上，并监测移植物的长期存活率和功能。使用 MMC 制造的细胞移植物具有低增殖率，同时仍允许与使用遗传药物选择获得的移植多巴胺神经元相比的稳健存活和功能。反过来，没有 FGF8 制造的细胞在移植后存活，但体内功能较差。我们的结果表明，MMC 可用于减少干细胞衍生的有丝分裂后神经元制剂中增殖细胞的数量，以用于 PD 细胞治疗。