3D spheroid formation and viability of
SGSCs under a priming (spheroid)
culture. (A) Floating, Matrigel, and microwell scaffolds induced SGSCs
to aggregate and assemble into 3D spheroids in a time-dependent manner.
Scale bars = 200 μm. (B) Diameter of spheroids was measured
after culture for 3 days, and the values were normalized to the total
number of spheroids. The data from five independent experiments were
analyzed and presented as a mean ± standard errors of the mean
(n = 5). One-way ANOVA, Tukey’s post hoc test.
*, compared with plastic; #, compared with floating culture; $, compared
to Matrigel. ***P < 0.001, ###P < 0.001, $$$P < 0.001.
(C) Spheroid-forming efficacy was determined through measurement of
the average number of spheroids per plate after plating the same number
of cells. The data from five independent experiments were analyzed
and presented as the mean ± standard errors of the mean (n = 5). One-way ANOVA, Tukey’s post hoc test. *,
compared with plastic; #, compared with floating culture; $, compared
with Matrigel. ***P < 0.001, ###P < 0.001, $$$P < 0.001.
(D) A representative LIVE/DEAD fluorescence image of hPECs spheroids
after culture for 5 days. (E) Cell viability percentage (viable cell
count/total cell count) was measured at 1, 3, 5, and 7 days by the
Trypan blue dye exclusion technique. The data from three independent
experiments were analyzed and presented as the mean ± standard
errors of the mean (n = 3).