Fig. 5
3D-primed salivary gland-resident stem cells (SGSCs) restore IR-induced salivary gland hypofunction in vivo. a Fluorescent in-situ hybridization (FISH) analysis. b Representative histological images of hematoxylin and eosin (H&E), Periodic acid Schiff (PAS), and Masson’s trichrome (MTC) staining from three groups at 12 weeks post IR. Scale bars represent 50 μm. A acinar, D duct. Arrow: fibrosis. c Body and gland weights measured at 16 weeks after treatment. d Salivary flow rate (SFR) calculated at 16 weeks. Changes in SFR after IR expressed as ratio of post-IR SFR to pre-IR SFR (mean ± SEM). Time to salivation (lag time (LT)) measured and ratios of post-IR LT to pre-IR LT presented. e Salivary amylase activity examined using assay kit and fold-changes in activity levels presented. f EGF content measured at each time point and average concentrations presented. Data presented as mean ± SEM. One-way ANOVA; Tukey’s post-hoc test. *Compared with Sham group; #compared with IR group; $compared with SGSC2D group. ***P < 0.001, #P < 0.05, ##P < 0.01, ###P < 0.001, $P < 0.05. g Differentially regulated genes (> 2-fold) quantified and presented in a heat map indicating antibody reactivity intensity as red (values > 2-fold) and green (values < 2-fold) pixels. h, i Salivary acinar and growth factor genes in SGSC3D group after whole-genome microarray analysis and comparison with IR and SGSC2D groups. One-way ANOVA; Tukey’s post-hoc test. *Compared with IR group; #compared with SGSC2D group. ***P < 0.001, ###P < 0.001. SGSCs2D monolayer-cultured SGSCs, SGSCs3D 3D spheroid-derived SGSCs, IR irradiation, EGF epidermal growth factor