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. 2017 Aug 10;9(3):807–819. doi: 10.1016/j.stemcr.2017.07.012

Figure 2.

Figure 2

Impact of Hypoxia and Nutrient Deprivation on Islets and SCIPC Cell Death

(A) SCIPC, human islets, and mouse islets were cultured at different densities for 6 hr as described in Experimental Procedures. Percentages of PI+ dead cells were quantified using flow cytometry. Data are a compilation of results from five independent experiments (SCIPC: n = 12, 10, 6; human islets: n = 6 at each density; mouse islets: n = 9 at each density).

(B) SCIPC and mouse islets were cultured in complete or diluted RPMI medium for 6 hr. Cell viability was quantified as described in (A). Data are a compilation of results from four independent experiments (SCIPC: n = 9 per condition; mouse islets: n = 4 per condition). For (A) and (B), statistical significance of the differences among each cell type at different densities is determined using one-way ANOVA with Holm-Sidak multiple comparisons.

(C) GSIS was measured using mouse islets after 6-hr low-density and high-density cultures. Data are a compilation of results from three independent experiments with paired low- and high-density cultured islets (n = 3 per group). Statistical difference was calculated using two-way ANOVA with Sidak's multiple comparisons test.

(D) Stimulation index of mouse islets shown in (C). A two-tailed paired t test was used to determine statistical significance of the difference (p = 0.0020).

(E) SCIPC, human islets, and mouse islets were cultured for 24 hr in the presence of 21% or 1% oxygen. At the end of the experiment cell viability was measured as described in (A). Data are a compilation of results from three independent experiments (n = 6–7 per condition for each cell type). Statistical significance of the difference between 21% and 1% oxygen for each cell type was calculated using an unpaired t test with Welch's correction.

(F) SCIPC, human islets, and mouse islets cultured for 24 hr at various densities with 1% or 21% oxygen. At the end of the experiment cell viability was measured as described in (A). Data are a compilation of results from three independent experiments (n = 6 per condition for each cell type). Statistical difference was calculated using one-way ANOVA with Holm-Sidak multiple comparisons.

All data are expressed as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001; ns, not significant.