Transcriptional Characterization of Human Myenteric Ganglia-Derived Neurospheres

Single-cell RNA sequencing (scRNA-seq) was performed to characterize the heterogeneity of enteric neural stem (ENSCs) cultures isolated from human intestinal samples. Dissection of the intestinal muscularis propria and enzymatic digestion was performed followed by the culture of either, 1) single cell suspensions, 2) counter filtered material >100 microns enriched for myenteric ganglia, or 3) hand-picked myenteric ganglia under a dissection microscope. Neurosphere cultures were characterized using the Chromium Single Cell 3' Reagent Kit v3.0, sequenced on an Illumina NextSeq platform with counts data generated using the 10X genomics Cell Ranger Count v7.1.0 pipeline. These data have been generated to understand the transcriptional landscape of ENSCs generated from human specimens and identify potential markers indicative of their neurogenic potential and functional properties.