GSM9779491: APOEε4-Vehivle brain organoids, day 60; Homo sapiens; RNA-Seq
For single-cell dissociation, three brain organoids per sample were pooled. The culture medium was removed, and the organoids were washed with 0.5 mM EDTA in PBS and incubated for 2 minutes at room temperature; this washing step was repeated once. Subsequently, the organoids were dissociated using ACCUMAX (Innovative Cell Technologies, San Diego, CA, USA). The samples were incubated at 37°C for 4 minutes, gently agitated, and incubated for an additional 4 minutes at 37°C. Mechanical dissociation was performed by pipetting until a single-cell suspension was achieved. To remove cell clumps and debris, the suspension was filtered through a 40 µm Flowmi cell strainer. The cells were centrifuged at 200 × g for 5 minutes at 4°C. The supernatant was discarded, and the cell pellet was washed and resuspended in PBS containing 0.04% BSA. The washing step was repeated to ensure high sample quality for library preparation. Each sample was loaded into one lane of a 10x Genomics microfluidic chip (Chromium Next GEM-X 3′ Chip) at a concentration adjusted to live cell counts, according to the manufacturer's instructions.
GEO
GSM9779491_r1