Human autologous iPSC–derived dopaminergic progenitors restore motor function in Parkinson’s disease models
Bin Song, … , Jeffrey S. Schweitzer, Kwang-Soo Kim
Bin Song, … , Jeffrey S. Schweitzer, Kwang-Soo Kim
Published November 12, 2019
Citation Information: J Clin Invest. 2020;130(2):904-920. https://doi.org/10.1172/JCI130767.
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Research Article Neuroscience Stem cells

Human autologous iPSC–derived dopaminergic progenitors restore motor function in Parkinson’s disease models

Abstract

Parkinson’s disease (PD) is a neurodegenerative disorder associated with loss of striatal dopamine, secondary to degeneration of midbrain dopamine (mDA) neurons in the substantia nigra, rendering cell transplantation a promising therapeutic strategy. To establish human induced pluripotent stem cell–based (hiPSC-based) autologous cell therapy, we report a platform of core techniques for the production of mDA progenitors as a safe and effective therapeutic product. First, by combining metabolism-regulating microRNAs with reprogramming factors, we developed a method to more efficiently generate clinical-grade iPSCs, as evidenced by genomic integrity and unbiased pluripotent potential. Second, we established a “spotting”-based in vitro differentiation methodology to generate functional and healthy mDA cells in a scalable manner. Third, we developed a chemical method that safely eliminates undifferentiated cells from the final product. Dopaminergic cells thus express high levels of characteristic mDA markers, produce and secrete dopamine, and exhibit electrophysiological features typical of mDA cells. Transplantation of these cells into rodent models of PD robustly restores motor function and reinnervates host brain, while showing no evidence of tumor formation or redistribution of the implanted cells. We propose that this platform is suitable for the successful implementation of human personalized autologous cell therapy for PD.

Authors

Bin Song, Young Cha, Sanghyeok Ko, Jeha Jeon, Nayeon Lee, Hyemyung Seo, Kyung-Joon Park, In-Hee Lee, Claudia Lopes, Melissa Feitosa, María José Luna, Jin Hyuk Jung, Jisun Kim, Dabin Hwang, Bruce M. Cohen, Martin H. Teicher, Pierre Leblanc, Bob S. Carter, Jeffrey H. Kordower, Vadim Y. Bolshakov, Sek Won Kong, Jeffrey S. Schweitzer, Kwang-Soo Kim

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Figure 5

Effects of quercetin treatment on undifferentiated and differentiated cells.

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Effects of quercetin treatment on undifferentiated and differentiated ce...
(A) Screening to determine optimal quercetin treatment conditions. Surviving hiPSCs were counted using a hemocytometer after treatment with different quercetin concentrations and durations. (B and C) Viability (B) and total cell number (C) of dopaminergic cells at d11 after quercetin treatment on d9. Cultures were treated for 16 hours at 5, 10, 20, 40, and 100 μM. Mean ± SD. n = 4. One-way ANOVA. (D) Colony formation by hiPSCs serially diluted by factors of 10 from 105 to 1 together with a constant number of fibroblasts (105). Cells were treated with 40 μM quercetin for 16 hours or left untreated, and then cultured for 6 days, followed by staining for AP activity. Representative results from 2 separate experiments. (E) Plotting of final colony number counted against original input hiPSC number. (F) Generation of standard curve for OCT4 copy number against input hiPSC number by qRT-PCR. OCT4 copy number was measured by qRT-PCR and calculated from 10-fold serially diluted hiPSCs, from 105 to 102 cells. (G) Measurement using OCT4 qRT-PCR of OCT4-positive cell numbers among mDA cells differentiated from hiPSCs at various time points with or without quercetin treatment. Mean ± SD. n = 2. ***P < 0.005, 2-way ANOVA.