Advanced patient-specific microglia cell models for pre-clinical studies in Alzheimers disease

We characterized monocyte-derived microglia (MDMi) cultured in 2D, 3D, and 3D co-cultured with ReNcell VM, with our data demonstrating that 3D co-culture closely resembles human brain microglia compared to 3D MDMi and 2D MDMi. The experimental design involved the collection of longitudinal blood samples from a single healthy male control donor under 40 years of age, free of dementia, and without a family history of Alzheimer's disease (AD). Blood collections were performed bi-monthly over a four-month period, resulting in multiple time points. Using EDTA tubes, peripheral blood mononuclear cells (PBMC) were isolated following the SepMate50 protocol. MDMi were cultured for 14 days (2D) or 35 days (3D) under a culture medium containing IL-34 and GM-CSF. 3D mono-culture and 3D co-culture were grown in thick Matrigel, while 2D was cultured on Matrigel-coated plates. For RNA collection, MDMi from all conditions were isolated and sorted for CD11b+ (thereby excluding leukocytes and ReNcell VM) and fixed according to the Chromium fixed RNA profiling workflow. Library construction involved simultaneous processing of all samples.