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. 2023 Jan 26;13:976945. doi: 10.3389/fonc.2023.976945

Figure 4.

Figure 4

Assessment of brain slice viability in culture. (A) Brain slices were collected from 6, 8, or 22 week old mice and cultured for 4 days in vitro (DIV). Cell viability was assessed by incubating brain slices in 10% alamarBlue™ for 24 hours. Autoclaved alamarBlue™ solution (100% reduced) is used as the positive control. Data are mean ± SD (n=4 mice, **p< 0.01, ***p < 0.001, one-way ANOVA with Tukey’s multiple comparison test). (B) Brain slices were collected in duplicate from 6-week old male mice and cultured in serum-free medium (SFM), medium containing 1X B27 supplement, DMEM containing 5% or 25% fetal bovine serum (FBS), or DMEM containing 5% or 25% heat-inactivated horse serum (HI-HS) for 4 days in vitro (DIV). Data are mean ± SD (n=3 mice, *p< 0.05, **p < 0.01, one-way ANOVA with Tukey’s multiple comparison test). (C) Brain slices collected from 6-week old mice were incubated in DMEM containing 5% FBS for 0, 2, 4, 7, and 14 days in vitro (DIV) and brain slice viability was assessed via alamarBlue™ intensity. Data represent the mean ± SD (n ≥ 2 mice). (D) Brain slice viability of each slice collected from 8-week old mice was determined via alamarBlue™ intensity at 0 DIV (n=2 mice). 10% alamarBlue™ solution with no brain slice represents the negative control.