Figure 1.
iPSCs from Control Subjects and MS Patients Efficiently Differentiate into Mature Astrocytes
Astrocytes were derived from seven donors (three healthy controls [HCs] and four MS). Two hiPSC lines were derived for each study subject except for HC1 (one hiPSC line).
(A) Timeline representing the different steps of hiPSC differentiation into mature astrocytes.
(B) Representative immunofluorescence staining images of hiPSC-derived astrocytes expressing GLAST (red), S100β (blue), and GFAP (green) obtained from healthy control HC2. Scale bar, 100 μm. Pictures were acquired with a Zeiss LSM 880 confocal microscope.
(C) Expression of the markers GLAST, S100β, and GFAP in hiPSC-derived astrocytes and primary astrocytes as assessed by flow cytometry. Bars represent the global mean ± SEM. Wilcoxon test was performed to assess statistical significance (∗p < 0.05). Each experiment was performed in duplicate and each dot represents the mean results of one or two experiments performed on a given cell line.
(D) Gene expression of 35 markers from hiPSC-derived astrocytes, primary astrocytes, hiPSCs, hiPSC-derived GPCs, and hiPSC-derived neurons measured by high-throughput qRT-PCR on the Biomark (Fluidigm). The 35 markers are listed on the x axis, and cell type and donors on the y axis. Results are expressed as the Z score of the –ΔCT (CT of gene of interest – CT of GAPDH).
(E) Principal-component analysis including all the 35 markers measured by high-throughput qRT-PCR on the Biomark (Fluidigm).
(F) Venn diagram of the genes highly expressed (LogCPM > 5) by resting human primary astrocytes and hiPSC-derived astrocytes based on RNA sequencing data.
NSCs, neural stem cells; AGPCs, astrocyte-committed GPCs; SB, SB431542; FGF2, fibroblast growth factor 2; EGF, epithelium growth factor; LIF, leukemia inhibitory factor; CNTF, ciliary neurotrophic factor; HC, healthy control; MS, MS patient; pA, primary astrocytes.