Single cell RNA Sequencing of term Umbilical cord blood mononuclear cells

In this study, we used thawed UCBMC (n=2 lean and n=2 obese) and enriched for live cells using SYTOX Blue (1:1000) on the BD FACA Aria Fusion into RPMI (supplemented with 30% FBS). Sorted cells were counted in triplicates and resuspsended in PBS with 0.04% BSA in a final concentration of 1200 cells/uL. Cells were immediately loaded in the 10X Genomics Chromium Controller with a loading target of 17,600 cells. Libraries were generated using V3 chemistry per manufacturer's instructions (10X Genomics). Libraries were sequenced on Illumina NovaSeq with a sequencing target of 50,000 reads per cell.