Gene expression profile at single cell level of hippocampal organoids

Hippocampal organoids are derived from human induced pluripotent stem cells under the guidance of multiple growth factors, such as proteins, chemical inhibitors, and activators. We used single-cell RNA sequencing (scRNA-seq) to analyze the transcriptomic signature of hippocampal organoids so that we could characterize cell identities in them. We prepared two single-cell samples from day-70 and day-81 hippocampal organoids. Overall design: We used 4 day-70 hippocampal organoids to prepare sample #1. Organoids were dissected into fragments and plated on flasks. We cultured them for 2-4 days, then isolated them with a Pierce Primary Neuron Isolation Kit with papain enzyme. We used 10 day-81 hippocampal organoids to prepare sample #2. Organoids were isolated directly with a Pierce Primary Neuron Isolation Kit with papain enzyme, broken up by pipetting up and down. The pre-sorting of live cells was completed using magnetic-activated cell sorting. The 3' Gene expression Library was prepared with the quality-qualified cDNA based on Chromium Next GEM Single-cell 3' GEM protocol. The quality of the library was assessed according to the standard protocol after fragmentation, adaptor ligation, and sample index PCR.