scRNAseq analysis of four populations sorted from midbrain organoids using Flow cytometry

Midbrain organoid derived from human induced pluriopotent stem cells a new model of Parkinson's Disease. Organ specific organoids can be used to model many diseases, however little is know about these models. We created a flow cytometry workflow to identify cell types in midbrain organoids. We then selected four populations and sorted these populations: Neurons1(CD24++), Neurons2(CD56++), Astrocytes and Radial Glia, using FACS and the antibody intensity gates defined by our workflow. We then performed scRNAseq on the sorted population and identified cell types within the sorted populations. Overall design: Human midbrain organoids dervived from the iPSC line AIW002-02 were cultured 304 days in final differenation media. Three samples of 20 organoids were discociated. The single cell suspensions were labelled with a panel of 13 different antibodies. Individual samples were hashtag labelled. Using the CelltypeR workflow gates for 4 populations were defined and cells were sorted using FACS. Technical replicates were pooled prior to single cell sequencing.