Single glomerular mesangial cells RNA-seq analysis

Available single-cell RNA-seq analyses have revealed that individual cells of the same type differ substantially in gene expression. We wonder whether glomerular mesangial cells, the cell type that support the unique structure of glomeruli, also exhibit big difference in gene expression among individual cells; and what biological information could be obtained from the single-mesangial cell RNA-seq data. Therefore, we isolated mouse glomeruli by Dynabead/magnetic concentration method, and digested them with enzymes to dissociate the cells. We loaded the single cell suspension to a Fluidigm C1 Single-Cell Auto Prep System for single cell cDNA preparation. The cDNA samples were analyzed with mesangial cell marker, Gata3,and the Gata3 positive samples were amplified and underwent sequecing using Illumina Highseq 2000 system. Overall design: In total, 14 single mesangial cells from a C57BL/6 male mouse were analyzed