The hippocampus is important for memory formation and is severely affected in the brain with Alzheimer's disease (AD). Since AD brain tissue is available postmortem, our understanding of early pathogenic processes occurring in hippocampi remains speculative. Here, an MS-based proteomic approach was used to analyze free-floating hippocampal spheroids (HSs) enriched in PROX1-positive granule neurons, from induced pluripotent stem cells (iPSCs) of healthy individuals and AD patients. HSs generated from two AD patients carrying variations in amyloid precursor protein (APP) or presenilin 1 (PS1) genes, and their age and gender matched controls.