Maternal Schistosomaisis Early bone marrow B cell scRNA CITEseq

We performed scRNA sequencing to find transcriptional differences in early B cell populations in the bone marrow of mice born to mothers chronically infected with Schistosoma mansoni. Cite-seq was used to help identify cluster types using surface protein markers. Overall design: Hematopoietic stem and progenitor cells and B cells were purified from bone marrow from the femur of mice born to S. mansoni infected and control mothers using a FACs Aria. Sorted cells were DAPI-CD3-CD4-CD8a-CD11b-CD11c-GR-1-Ter119-CD19-. CD19 was used in place of B220 to enrich for pre-pro B cells, but the antibody was ineffective, allowing for all B cell types to be sorted. After sorting, cells were stained with the cell surface proteins CD117(cKit) and Sca-1 with oligonucleotide-tagged antibodies to perform CITE-sequencing. Bone marrow from at least 3 mice were pooled before sorting. Cells were loaded into the 10X Genomics Chromium instrument according to the manufacturer's directions.