Profiling Normal Bone Marrow B-Cell Subsets

Prior to study commencement, the Health Research Ethics Committee for the North Denmark Region approved our study protocol (MSCNET, N-20080062MCH). Following informed written consent, obtained in accordance with the Declaration of Helsinki, normal BM was collected from the sternum of seven adult patients undergoing cardiac surgery as described in the S1 Appendix, and elsewhere [12 (link)]. Fluorescence-activated cell sorting (FACS) was used to fractionate mononuclear BM cells into six distinct normal B-cell subsets: pre-BI (BI), pre-BII (BII), immature (IM), naïve (NA), memory (ME), and plasma cells (PC) (S1 Appendix and S1 Table). For gene expression profiling (GEP), mRNA from B-cell subsets were isolated and hybridized to the Human Exon 1.0 ST (Exon) array platform [12 (link)]^. A total of 38 CEL files containing B-cell data were generated using the Affymetrix Command Console. The CEL files and metadata were adjusted for compliance (with the requirements of Minimum Information About a Microarray Experiment) [16 (link)], and then deposited in the NCBI Gene Expression Omnibus repository (accession code GSE68878). In the present study, these data are referred to as the sternal BM B-cell data.