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. 2020 Sep 2;10:14474. doi: 10.1038/s41598-020-71329-8

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Differentiation and characterization of hiPSC-derived astrocytes from healthy subjects and PD patients. (A) Representative images from astrocyte differentiation. Bright field images showing hiPSCs at day 1, the rosettes at day 16, progenitor cells expanded as spheres from day 18 onwards and matured astrocytes at day 180. (B) Representative fluorescence images of astrocytes matured for 7 days and stained for AQP4, Vimentin, GFAP and S100B. Nuclei are stained with DAPI. Scale bars, 50 µm. (C) Relative gene expression levels GFAP, SLC2A1, SLC1A3 and AQP4 in astrocytes shown as fold change from healthy control. (D) Representative FACS histogram of glucose uptake analysed by fluorescent glucose analog 2-NBDG. Grey area shows untreated cells, black line shows cells incubated with 2-NBDG and red line shows cells incubated with insulin and 2-NBDG. (E) Glucose uptake analyzed by fluorescent glucose analog 2-NBDG. (F) Glutamate uptake measured with radiolabeled glutamic acid. Three independent experiments. Bars represents mean ± SD, *p < 0.05. PD1 patient carries a mutation in LRRK2 (G2019S) and GBA (N370S), PD2 patient carries a mutation in LRRK2 (G2019S) and isogenic control for PD2 patient.