Figure 5.

SB-HEF-iAs display ATP-induced calcium response and are immunocompetent

(A) Background subtracted images of live-cell calcium imaging experiments on Fluo-4-loaded rtTA-HEFs and SB-HEF-iAs at 5 weeks, and primary human fetal astrocytes (hAs).

(B) Mean relative fluorescence changes of calcium traces of ATP-responding cells normalized to baseline (dF/F0).

(C) Percentage of ATP-responding cells using a threshold of 3 × SD of baseline.

(D) Percentage of ATP-responding cells of S100B, GFAP, and S100B/GFAP co-expressing SB-HEF-iAs.

(E) Percentage of oscillating cells of ATP-responding cells.

(F) Amplitude of ATP-evoked response in responding cells.

(G and H) qRT-PCR analysis upon stimulation with TNFα, IL-1α, and C1q. Data are presented as fold change relative to non-stimulated controls.

(I) Representative immunostaining and bright-field image of SB-HEF-iAs following treatment with TNFα, IL-1α, and C1q.

Data are presented as mean ± SEM, except for calcium traces (B) presented as mean ± SD, of n = 3–4 independent experiments. Kruskal-Wallis test (C–F) and Mann-Whitney test (G and H) were performed for statistical analysis. ^∗p < 0.05, ^∗∗∗p < 0.001. Scale bars, 50 μm (A and I). See also Figure S6 and Video S1. Calcium imaging of rtTA-HEFs, Video S2. Calcium imaging of SB-HEF-iAs, Video S3. Calcium imaging of primary human fetal astrocytes.