Method Profile
Survival, lipid, mitochondrial, expression and calcium-imaging readouts compare semaglutide responses in forebrain organoids at different maturation stages.
Mechanistic and functional comparison of drug responses under neural metabolic stress.
Forebrain organoids derived from DXR0109B and DYR0100 hiPSC lines, with complementary two-dimensional neural progenitor cell assays.
Early and day-65-to-80 exposure windows; 100 nM semaglutide, 10 μg/mL cholesterol and corresponding unstressed, stressed and vehicle controls.
Human forebrain models
hiPSC-derived forebrain organoids and neural progenitor cells; the organoids lack microglia, functional vasculature, aging-related features and Alzheimer’s-associated genetic backgrounds.
Pharmacological comparisons
The GLP-1 receptor antagonist C5d and PKA/PI3K inhibitors probe receptor dependence and signaling contributions.
Functional and molecular readouts
Live/dead imaging, neural progenitor cell CCK-8 assays, lipid droplets, reactive oxygen species, protein and RNA measurements, and Fluo-4 AM calcium imaging analyzed with Fiji and Python.
Compare treatment and pharmacological blockade within each maturation stage. Interpret transcript-based cell–cell communication as an inference and Fluo-4 signals as cellular calcium activity.
Cholesterol stress with semaglutide co-treatment, receptor blockade and separate early- and later-stage assessments.
Early Figure 2 size measurements used 13 organoids per group, viability assays six independent experiments and protein measurements four. Sample sizes differ by endpoint.
Unstressed controls, cholesterol alone, cholesterol with semaglutide, and pharmacological blockade conditions.
Early survival, size and apoptosis measures improved. At day 80, overall area and APP/p-tau217 did not significantly recover; calcium event frequency improved while synchrony recovery was limited.
Semaglutide partly protected human forebrain organoids against the structural and metabolic effects of cholesterol stress. The response was more limited in older cultures, where some calcium activity measures improved without restoring tissue size or several disease-associated markers.
