GSM8751304: scRNA_seq_donor2_V5_TCR; Homo sapiens; RNA-Seq
Human breast milk samples were collected (between 30 to 50 ml) from donors in the morning on the day of experiment and were processed right away to retain high viability of the cells. The expressed milk samples were diluted with a similar volume of PBS before being centrifuged at 400g for 10min at 4C. After centrifugation, the top layer of fat and the supernatant were discarded. Cell pellets were resuspended in 50ml of PBS and underwent centrifugation again at 1350rpm for 5mins at 4C. Next, the cells were stained with human CD3 MicroBeads (Miltenyi Biotec) and enriched for T cells using the LS columns (Miltenyi Biotec). 8,300 – 10,000 cells from the enriched fraction were loaded to a Chromium Single Cell Controller (10X Genomics) to encapsulate cells into droplets. Libraries were prepared using a Chromium Single Cell 5' Reagent kits following manufacturer's instructions. Libraries were then sequenced on Illumina NovaSeq 6000 (Illumina). scRNA-seq