GSM8427621: Tonsil; Homo sapiens; RNA-Seq
A human tonsil specimen obtained by surgical resection was mechanically processed and cells were preserved frozen. Cells were later thawed and stained with the following flow cytometry panel: anti human CD45RA-FITC (HI100), anti human HLA -PercpCy5.5 (L243), anti human TCRab-BUV737 (IP26) and anti human IgD-BUV395 (IA6-2) antibodies from BD: anti human CD69-PECy7 (FN50), anti human CD45RO-BV605 (UCHL1), anti human CD8-BV421 (SK1), anti human CD4-BV786 (SK3), anti human CD38-APCCy7 (HIT2) and anti human CD19-AF700 (HIB19) antibodies from Biolegend. Next, cells were FACS sorted as follows using an Aria lll sorter (BD) and the indicated number of cells from each subset were combined in preparation for one 5' 10x genomics scRNAseq capture.   T cell sample: CD45RA+CD45RO+ CD4+ T Cells (50K), HLADR+CD38+/- CD4 T Cells (20K) and total TCRab+ CD4+ cells (10K).  B cell sample: TCRab- CD4- CD8- (20K), TCRab- CD4- CD8- IgD- (20K), TCRab- CD4- CD8- IgD- CD38+ (20K), TCRab- CD4- CD8- IgD- CD38+++ (15K). Library was constructed using the 10x Genomics 5' v2 kit according to the manufacturer's instructions.